CN100497580C - Cell handling device, tissue regeneration composition, and tissue regeneration method - Google Patents

Abstract

A cell handling device capable of storing and carrying sampled or cultured cells without being contaminated from the surroundings in regeneration treatment and easily injecting the cells in the living body, wherein the cells are sampled from the living body, the sampled cells or the cells provided by culturing the cells are stored in a syringe type storage container, and the cells stored in the storage container are transplanted into the living body. The cell storage container is desirably such that a part or more of the inner wall of the container in contact with the cells is formed with a cell unbonded material. Since the cells can take in oxygen required for the living thereof in the container and are prevented from being adhered to the inner wall of the container, the cells can be easily and rapidly transplanted to the living body without applying cell releasing treatment to the cells when the cells are used in the field of the regeneration treatment.

Description

Cell handling device, tissue regeneration are with composition and tissue regeneration method

Technical field

The present invention relates to regenerative medicine, be particularly related to the cell handling device and the tissue regeneration composition that are used for regenerative medicine.

Background technology

In recent years, along with the progress of molecular cytobiology, cellular engineering, the differentiation that will from organism, take out cell or cell (stem cell) with many differentiation functions and the of self-replication capacity transplant in the patient also carrying out always with the research of the regenerative medicine for the treatment of.So-called regenerative medicine is meant the methods of treatment that the cell that will break up or stem cell transplantation make destination organization (containing internal organs) repair and recover to the responsibility latibulum position of the defect of bio-tissue or disease.In general, in regenerative medicine that the cell of gathering is separately temporary transient or cultivate in special container in the presence of support, with the differentiation that obtains thus cell or undifferentiated cell transplant in target site by Surgery Treatment.

Like this, in regenerative medicine, be accompanied by the otomy that is used for Transplanted cells, often carry out Surgery Treatment, big to patient's burden, therefore study the cell that will cultivate and directly be injected into intravital method.As injecting chondrocyte or its precursor cell to the joint, injecting neurocyte, physiologically active substance generation cell or its precursor cell, inject myocardial cell or its precursor cell etc., can become strong methods of treatment to heart to brain.After the operations such as when implementing this treatment, the cell that process uses trypsinase, EDTA (ethylenediamine tetraacetic acid (EDTA), common name edetic acid) etc. will adhere to wall of container is peeled off, washing, gather specified amount.Usually use syringe or conduit to inject in the organism.

In addition, in the Transplanted cells of regenerative medicine, the not only collection of cell, cultivation, induction, a series of operation such as to transplant in organism cumbersome, and for prevent from around pollution (being infected with), must and use skilled sophisticated technology to carry out a series of operation under the environment of cleaning.Therefore, if the personnel that do not have the complete facility of equipment, have sophisticated technology are difficult to carry out cellular transplantation therapy.In addition, except operation, as situation about why not being polluted by surrounding environment under, the cell delivery of having gathered or having cultivated is also had difficulty in specified devices such as sterilisable chamber or incubator.

Summary of the invention

The present invention finishes based on this background, and its main purpose is, in regenerative medicine, the cell that can will gather or cultivate under not by the situation of ambient contamination is preserved and carried, and can easily this cell be injected in the organism.

Yet in order to reach in this purpose research, known have the following listed problem of enumerating, so be purpose to solve these problems also among the present invention.

Promptly, no matter who in regenerative medicine in the past, does not exist can both carry out the cell handling device of regenerative medicine sequence of operations simply, above-mentioned sequence of operations is meant cell collection, preservation, cultivation, in use can be simply with this Transplanted cells in body.

Specifically, can not survive and breed if most cells does not adhere on the support, so cell adhesion is in the wall of the culture vessel that becomes support.Therefore, when cell being carried out the organism transplanting, the cell that must will adhere to vessel surface is peeled off.Can carry out physics to this cell and peel off, also can use medicaments such as trypsinase or EDTA to peel off, but these operations might produce bad influence by pair cell.And, if the personnel that this cell manipulation does not have the complete facility of equipment and has sophisticated technology are difficult to carry out.

Like this, we can say under the present case,, therefore can not carry out Transplanted cells simply owing to the cell of having cultivated temporarily must be stripped down from the wall of container face.

Therefore, the object of the present invention is to provide a kind of cell handling device, this device is compared with the past to have simple structure and can prevent to pollute and preserve cell well and in cellular transplantation therapy, need not carry out the processing of from container cell being peeled off, just can easily this cell be injected into organism.

Above-mentioned main purpose can be reached by the following method, promptly in the tissue regeneration method, the cell that to gather from organism or with the cell that obtains after this cell cultures be stored in as the syringe-type device of cell handling device (promptly, mechanical means such as the manual operation by piston, control air pressure are installed the application of force to this, the volume in the fluid storage space in this device is changed, make content inflow or outflow (by pressurization thus, content is discharged from relief outlet) device), and the Transplanted cells that will be stored in this device is in organism.

That is, by the cell handling device of use injector type, when carrying out Transplanted cells at the regenerative medicine scene, by the operation same with common medical syringe, can be in organism with Transplanted cells.Therefore, be not limited to have the personnel of tip and knack, can both be than being easier to and promptly carrying out Transplanted cells, therefore favourable.In addition, the cell handling device of injector type has output, the influx that can control content simply, and the advantage of pressing pace of change in the control.

In addition, also carried out relating to the invention of following cell handling device and tissue regeneration usefulness composition in order to reach above-mentioned purpose.With these invention combinations, then cultivate under can be not by the situation of ambient contamination, preserve, carry cell reaching, easily cell being injected into also is very effective on this main purpose in the organism.

In order to reach this purpose, the inventor etc. will make the cell handling device that can liquid thickly stores the mobile process object thing that contains cell as cellular processor tool of the present invention, its constitute at least with at least a portion at above-mentioned cells contacting position in be provided with the zone that makes that the gas (oxygen that is used to breathe, be used to carbonic acid gas of keeping pH etc.) of cells survival institute necessary amounts sees through.

Here, " make the zone (breathable zones) of gas permeation " and be meant the place that can contact by having ventilation property and being the formed zone of material of non-perviousness to liquid with cell.In the time can not breathable zones being set with the place of cells contacting, this breathable zones not necessarily must have the close property of liquid.

Breathable zones among the present invention is meant that the oxygen permeability is more than or equal to 0.1mL/cm ²The zone of 24hratm.Area for breathable zones is not particularly limited, but from the aspect of essential oxygen fully is provided to cell, the preferred cell treatment unit is more than or equal to 1mL/24hratm at the total oxygen permeating amount with the cell suspending liquid contact part, is preferably more than especially to equal 10mL/24hratm.

Like this, in cell handling device of the present invention, by breathable zones gas can circulate (gaseous interchange), therefore in the part outside this breathable zones, even if use close property of liquid and bubble-tight material, also can install inner liquid and thickly preserve cell suspending liquid, the picked-up by the necessary oxygen of cells survival, the gas regulation such as discharge of carbonic acid gas simultaneously, the keeping of states such as pH condition of realizing containing the nutrient solution of cell suspending liquid at this.Therefore, can in the inactivation of cell in this device of the collection that prevents to be used for regenerative medicine, carry this device, also can install inside simultaneously and make cell proliferation or induction well at this.

Here, if with the whole cell storage compartment that is arranged in the above-mentioned cell handling device of above-mentioned breathable zones,, easily impartial and bring into play gaseous interchange fully then with respect to the cell integral body in the cell suspending liquid.Therefore, even if be not so excellent poromeric material, also can access sufficient gaseous interchange.

On the other hand, use when having the material of more excellent ventilation property, there is no need to constitute the whole of above-mentioned cell handling device, need only being provided with in the cell storage compartment at least with dividing with the contacted upper inside wall of cell suspending liquid with this material.At this moment, can form zone with shapes such as rectangle, circles with regulation area.

Under the situation of said syringe type cell handling device, preferably on the part of the piston of the whole or part of the main part that stores cell, exposing cell, form breathable zones.When on the part of aforementioned body portion, forming breathable zones, because area is limited, therefore also depend on the essential minimum concentration of cells survival desired gas (oxygen, carbonic acid gas), the preferred higher material of ventilation property that uses designs according to the mode of the required gas of can fully guaranteeing to survive.In other words, the amount of cells survival desired gas is different and different according to cell, but in order to make cells survival, the preferred material with highly air-permeable that uses carries out gaseous interchange fully.

In addition, owing to reasons such as the range of choice of material limit to some extent, when the part of aforementioned body portion forms breathable zones, by forming independently a plurality of separately zones, then impartial and abundant amount ground increases to the effect of the whole supply gas of above-mentioned cell storage compartment, and is therefore preferred.

When the part of aforementioned body portion is provided with breathable zones, can list porous film as example with excellent breathability.In this porous film, control the aperture, can keep the close property of liquid thus, therefore find, if use the film that can guarantee fluid-tight degree is processed in the hole, even if smaller area also can be guaranteed sufficient Air permenbility.

In addition,, can also pass through porous film, existing bubble in the cell suspending liquid that is stored in the cell handling device is expelled to safely the outside of cell handling device if adopt porous film as above-mentioned breathable zones.

Under the situation of injector type cell handling device, the form as the main part that stores cell can list the form that directly cell is stored in cylindrical container (outer cylinder body); And will be accommodated in the intravital form of urceolus by so-called bag-like containers such as the reducible snake shape in internal space, cryptomere, tubulose by the effect of thrust pressure.Under the former situation, preferably at outer cylinder body, be used to guarantee that fluid-tight lid, piston etc. locate to form breathable zones.Under the situation of latter's form, by at least a portion that is accommodated in inner container, forming breathable zones, can be to the cell supply gas, and especially under latter instance, in outer cylinder body, take under the state of bag-like container, preferably according in the bag-like container and the device part (positions such as piston, outer cylinder body) of mode beyond bag-like container that can carry out gaseous interchange between the outside go up and form breathable zones.In addition, the container of bag shape loads and unloads freely, and the preservation of cell and cultivation are certain under the state of installation, also can implement the preservation and the cultivation of cell under the state of unloading.Its reason is that this container possesses ventilation property simultaneously originally as the close property of liquid.

In addition, as the film that forms breathable zones, the film that can use the macromolecular material by good air permeability to form, even if but ventilation property is not the macromolecular material of excellence so, by making porous film, and suitably setting the aperture, also can give ventilation property and to the non-perviousness of liquid.Therefore, making that cell handling device is the fluid-tight while, can prevent that also the part of cell suspending liquid from partly spilling from air-permeable envelope.

In addition, in the present invention, the contacted position of cell suspending liquid of using cell to be difficult to adherent material formation and cell handling device is effective.As the evaluation method of cell adhesion, can form the auxiliary protein that sticks together spot (adhesion plaque) by the immunological method detection and estimate, or estimate by calculating the cell adhesion number.

Like this, by reducing the cell adhesion of cell handling device inner face, can prevent in preservation that cell adhesion is in the cell handling device wall.Thus, can access following effect at the scene of regenerative medicine.

Promptly, all the time, the cell of in regenerative medicine, transplanting, as utilize cell cultures to wait with culture dish and cultivate preservation, but because cell adhesion must carry out the lift-off processing of cell in its inner face when therefore transplanting in organism in this cell handling device in the past.In addition,, must in highly antipollution equipment, operate, can not easily carry out regenerative medicine by skilled personnel with high technology in order to carry out this lift-off processing.Relative therewith, since cell handling device of the present invention with cell when this cell handling device takes out, the lift-off processing that does not need cell, do not carry out the strip operation of physics, do not make with medicament, can under the situation of not damaging cell, transplant, therefore can have the effect that to implement regenerative medicine well in organism.And because the lift-off processing of not carrying out cell just can will be stored in cell directly transplanting in the cell handling device in organism, therefore can save the loaded down with trivial details of graft procedure, the personnel of high degree of skill technology also can easily carry out the Transplanted cells operation even if do not have so simultaneously.

And, the inventor etc. are to the material and the shape of the support of the cell proliferation that is used to regenerative medicine is used, induction, research has repeatedly been carried out on the basis that is contemplated that with its uniqueness, found that the shape of support is made particle shape, used bioabsorbable material to form support simultaneously, can simplify the operation that is accompanied by cell cultures significantly thus, and then finish the research of tissue regeneration of the present invention with composition.That is, tissue regeneration composition of the present invention has mobile medium and the cytoskeleton particle that becomes cytoskeletal particle shape, and its formation is: the cytoskeleton particle is made of bioabsorbable material, and cell adhesion is on this cytoskeleton particle.

By this tissue regeneration composition of the present invention, can make the cell of gathering from organism in the surface of scaffold particle propagation or to the target cell induction.During this period, cell adhesion is bred in scaffold particle, has flowability usually in mobile medium (nutrient solution (comprising body fluid)).And, when carrying out Transplanted cells, this cell cultures particle directly can be injected in the organism.

Therefore, needn't as in the past, after cell proliferation, implement the cell lift-off processing, and can improve the efficient of the Transplanted cells operation in the regenerative medicine greatly.In addition, owing to needn't carry out the cell lift-off processing, therefore needn't worry damaging cells.This tissue regeneration when affected part injects, is not to use the material that is stained with cell on scaffold particle with composition, can only scaffold particle be injected yet, and becomes the support of host cell.

In addition, owing to use bioabsorbable material to form scaffold particle, therefore be injected into behind the organism through behind the certain hour, scaffold particle absorbs by organism and disappears.Therefore, the operation of removing support after also not needing to perform the operation.

Here, when cell was transplanted to human body, the method that the material that will be suspended with cell in nutrient solution is transplanted to human body was as a kind of trial, and in this method, because mobile high, so cell runs off, and cell is difficult to suitably fixed growth at transplantation site.For head it off, trial will be in disperseing with matrix (high-viscous solution, as gelatin, osso-albumin etc.), and the suspension (high viscosity solution) that disperses transplanting to obtain with cell is transplanted to the method for affected part, so that cell does not run off, but in this method, disperse the matrix of usefulness to become obstacle, thus suitably fixed growth on transplantation site.But, above-mentioned tissue regeneration composition, because cell adhesion in particle surface, when therefore attempting Transplanted cells, is difficult for this problem takes place, cell is fixed growth well.

And, because support is the particle shape, so the surface-area height of per unit volume, so on a spot of support, can adhere to a lot of cells effectively.

If the tissue regeneration that will be made of this cell cultures particle is kept in the above-mentioned cell handling device with syringe styled with composition, then when transplanting, can from this cell handling device, easily and promptly cell be transplanted in the organism.Therefore, can alleviate operation of patients such as for example physical relatively poor child, old man etc., and can in regenerative medicine, alleviate patient's burden.

And then, by this tissue regeneration is accommodated in the above-mentioned cell handling device with composition, can reach with the conveying of cell, cultivation, to the interim operation till the Transplanted cells of human body as a link, in a container the simple effect of implementing.Promptly, because above-mentioned cell handling device possesses breathable zones, therefore can in this device, preserve and cultivate, so can save specially with the operation of cell transfer in the cell cultures special container, and this device is the form of injector type, given play to thus and can use same apparatus, will preserve in inside and cultured cells is directly carried out the higher effect of graft procedure by conduits such as pin, conduits.

When utilizing above-mentioned cell handling device to implement to transplant, if cell adhesion is in installing wall, the cell count of then being injected reduces, and therefore the cell of necessary amounts can not be transplanted in the human body.Here, for head it off, it is effectively that the applicator inner-wall surface reduces the cell adhesion number with the NA above-mentioned characteristic of cell, and still, when using cultivation and differentiation phase to need the adherent cell of support, this cell has lost support.If it is use above-mentioned bioabsorbable scaffold particle this moment,, therefore very effective then for this cell provides support.

When cell adhesion position when being a plurality of, the adherent tendency in oriented easy adherent position, so scaffold particle compares with the device inner-wall surface in order to play a role, and must be the easier adherent material of cell certainly.

Description of drawings

Fig. 1 is the structure iron as the syringe of the cell handling device of embodiment 1.

Fig. 2 is the structure iron as the syringe of the cell handling device of embodiment 2.

Fig. 3 is the structure iron as the syringe of the cell handling device of embodiment 3.

Fig. 4 is the structure iron as the syringe of the cell handling device of embodiment 4.

Fig. 5 is the performance data of cell handling device of the present invention.

Fig. 6 is the structure iron of the cell handling device of embodiment 5.

Fig. 7 is the structure iron of the cell handling device of embodiment 6.

Fig. 8 is the structure iron as the syringe of the cell handling device of embodiment 7.

Fig. 9 is the structure iron as the syringe of the cell handling device of embodiment 8.

Figure 10 is the pie graph of tissue regeneration of the present invention with composition.

Embodiment

At first, tissue regeneration method of the present invention is described.

Tissue regeneration method of the present invention contains following step basically.

I. gather the acquisition step of cell from organism

At first, gather specific cell from organism.

Ii. separate and purification step

The cell of gathering by FACS separation such as (flow cytometers) and purifying.

Iii. cell is stored in the preservation step in the cell handling device

Cell be stored in cell handling device thereafter.

Iv. breed step

The cell handling device of preserving cell is stored in cell cultivation equipment (CPC) etc.Then, in this cell cultivation equipment etc., make the cell proliferation in the cell handling device and carry out induction as required.

V. transplant step

Use above-mentioned cell handling device, with propagation and induction Transplanted cells in organism.

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Here, in the step of iii～v, use the injector type cellular processor tool shown in the following embodiment 1～8, can in whole operations of the regenerative medicine of preserving, carry, breed, transplanting, use same device to carry out a series of operation thus, therefore can be not with cell transfer to other containers, can be safely and be advanced to next operation rapidly, making rapidly, treatment becomes possibility.

Particularly during the Transplanted cells in the regenerative medicine scene, its availability height can similarly be operated with common medical syringe, and the cell of preserving is transplanted in the organism.Therefore, be not limited only to have the personnel of height and knack, also can be easily and promptly carry out Transplanted cells.And, can also when preserving cell, carry.

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In addition, in the regenerative medicine scene of reality, mostly the place of carrying out each step is separated, but cell handling device of the present invention can handle hydraulic seal, therefore also is suitable for cell delivery to the place that separates.

For example, in the preservation step of iii, by be used for from organism cell is fetched into till the cultivation or till transplant during in cell preserve, also can be equally easily and promptly handle cell.Perhaps, in the propagation step of iv, also can use cell handling device of the present invention during culturing cell.And, can use as the culture apparatus of cell, can utilize as the means that former state is preserved cell thereafter.

In general, cell suspending liquid is by nutrient solution and cellularity.Nutrient solution can directly use the nutrient solution that in the past used, but the material (virus, prion etc.) that interpolation may become infection reason is not preferably added or reduces in the security when considering nutrient solution in cell is transplanted to organism as far as possible.

Here, in cell suspending liquid, can contain various cells in therepic use of being useful on regenerative medicine etc.Cell can use above-mentioned various cell category according to therapeutic purpose.This cell category is not particularly limited, concrete example is except listing embryo's property stem cell (embryonic stemcells:ES cell), embryo's sexual reproductive cell (embryonic germ cells:EG cell), adult stem cell (adult stem cells:AS cell, be also referred to as adult stem), mescenchymal stem cell, neural stem cell, the blood vessel endothelium stem cell, the hematopoiesis lineage stem cells, stem cells such as liver stem cells also can list osteocyte, the chondrocyte, the myocyte, the myocardial cell, neurocyte, tendon cell, adipocyte, pancreatic cell, liver cell, nephrocyte, hair follicle cell, cell or its precursor cell that hemocyte etc. have broken up.Like this, can use with each differentiation degree headed by embryo's property stem cell different stem cell and the cell that is divided into each tissue.Wherein, when using adherent cell, can effectively give the support of cell handling device with cell non-adhesive and use finely particulate.Reason is that adherent cell is at propagation and the essential support of differentiation phase.

This cell can be gathered by known method.That is,, after wherein the cell selective ground of necessity being separated, add growth factor or somatomedin etc. as required and cultivate from the regulation position chorista (cell) of organism.This cultivation is preferably implemented in the incubator of special use.Cultured cells thus is stored in the cell handling device inside of injector type, under the condition that is suitable for preserving, preserves until being used for the treatment of.

Provide the combination example of cell and nutrient solution.

Promptly, during culturing cell end user mescenchymal stem cell (human mesenchymal stem cell), can in the human mesenchymal stem cell basic medium (the POIETICS company U.S.) of 440mL, add growth of mesenchymal stem cells additive (50mL), L-glutaminate (10mL), penicillin/streptomycin (0.5mL) as nutrient solution.

Cartilage induction substratum (nutrient solution) when using the chondrocyte can use the substratum that has added dexamethasone (1mL), Sodium.alpha.-ketopropionate (2mL), xitix (2mL), proline(Pro) (2mL), ITS+ additive (2mL), L-glutaminate (4mL), penicillin/streptomycin (2mL) in the basic medium (185mL) of hMSC differentiation.

On the other hand, the tissue regeneration that is made of cell cultures particle of the present invention is described in detail in the back with composition.

Below, the situation of the cell handling device that uses injector type being preserved cell suspending liquid is elaborated.

1. about cell handling device of the present invention

(embodiment 1)

1-1. the structure of injector type cell handling device 1

Figure 1 shows that structure iron as the injector type cell handling device 1 of the embodiment 1 of a routine cell handling device of the present invention.Fig. 1 (A) is that stereographic map, Fig. 1 (B) are the X-X ' sectional view of Fig. 1 (B) for side elevational view, Fig. 1 (C).

The structure of injector type cell handling device 1 shown in Figure 1 roughly is divided into syringe body portion 2, piston (plunger) (being also referred to as thrust piece or piston) 40 etc.Store cell suspending liquid 100 in the syringe body portion 2.

Syringe body portion 2 is by being that cylindrical body 3 and the air-permeable envelope described later 20 that drum forms constitutes with cell non-adhesive material injection forming.

The front end side of cylindrical body 3 is formed with discoid front end face 110 and outstanding bait portion 120 thereon.The front end of bait portion 120 has been executed usually and has been covered 60.Insert piston 40 from rear end 12 sides of syringe body portion 2, syringe body portion 2 inside are sealed as the close property of liquid thus.In addition, remaining inside of bait portion 120 seals with resin etc., and (during Transplanted cells) can be with seal break-off in use.

The material of the cylindrical body 3 here can use the material that is generally used in the syringe so long as the material that can be shaped gets final product, but as one of feature of the present invention, the material of preferred cylindrical body 3 uses the material that is difficult to adherent cell.Detailed content as described later, the part of cylindrical body 3 is used this cell non-adhesive material, can prevent cell adhesion thus in cylindrical body 3 inside, and good preservation in can be in being suspended in nutrient solution.

Here said " cell non-adhesive " is even if be that phalangeal cell does not adhere to or adhere to the degree that can simply peel off that also just adhered to fully.

Piston 40 is by forming material injection formings such as polyethylene, polypropylene, polycarbonate, polyvinyl chloride, has the structure that is formed with at the two ends of the main body 42 of cruciform cross-section shape in the discoid end of diametric(al) expansion.One in the above-mentioned end is the piston crown 43 that is inserted into syringe body portion 2 inside on direction of principal axis, the pushing end 41 that the other end pushes with finger for the user, and this pushing end 41 is used for piston 40 is pushed into syringe body portion 2 inside.

Piston head 43 is made of elastomeric element, makes in syringe body portion 2 (being cylindrical body 3 and air-permeable envelope 20 in detail) inside, can thickly keep cell suspending liquid 100 by liquid.

(about cell non-adhesive material)

Here " whether cell is difficult to adherent evaluation ", " evaluation that whether has the cell non-adhesive " in other words, estimate as forming the auxiliary protein that sticks together spot (adhesion plaque), or estimate by calculating the cell adhesion number by the immunological method detection.

As above-mentioned cell non-adhesive material, preferable material is as follows.That is, according to the difference of the cell category of preserving and difference, but in general, the material and the surperficial material that has negative charge that are formed by hydrophilic material or hydrophobic material have the cell non-adhesive more, can be used as preferable material.As hydrophilic material, be 50 degree or following materials preferably to water contact angle; As hydrophobic material, be 100 degree or above materials preferably to water contact angle.

As the example of preferred hydrophilic material, can list substrate surface by methods such as graft copolymerization or chemical reaction in conjunction with or be coated with the material of acrylamide polymer, Methacrylamide based polymer, polyacrylic acid, polyvinyl alcohol, polyoxyethylene glycol, polyvinylpyrrolidone, Mierocrystalline cellulose, dextran, hyaluronic acid, glucosaminoglycan, protein-polysaccharide, carrageenan and protein etc. from the teeth outwards.About the adjustment of substrate surface with cover and handle, must after utilizing making base material such as injection forming, carry out in addition.

As hydrophobic material, can list fluorine resin and silicone resin such as tetrafluoroethylene, tetrafluoraoethylene-hexafluoropropylene copolymer, polyethylene terephthalate, polypropylene.

Have the material of negative charge as the surface, can list the material that is combined with polyacrylic acid, polymethyl acrylic acid, styrene sulfonic acid, alginic acid, heparin, Suleparoid, chondroitin sulfate or dermatan sulfate from the teeth outwards.Wherein special preferred surface has the material of carboxyl.The surface smoothing of this material, excellent on the cell non-adhesive, therefore preferred.

Because silicone resin is also excellent on ventilation property, so it is a preferable material in the above-mentioned materials.In addition,, can access higher ventilation property because tetrafluoroethylene and tetrafluoraoethylene-hexafluoropropylene copolymer are compared with porous film, therefore same preferred.

Preferred this cell non-adhesive material that uses constitutes and the contacted packing ring part of cell.

(about breathable zones)

The perisporium 30 of cylindrical body 3 is provided with the communicating pores that connects cylindrical body 3 on thickness direction.Here shown in Fig. 1 (A), for a plurality of (being 4 here) with the OBL slit 31 of this syringe direction of principal axis as length direction, but the example that this OBL slit 31 is the communicating pores shape, its shape and number are not limited thereto.

And, shown in Fig. 1 (C), be provided with air-permeable envelope 20 cylindraceous in slit 31 inside, this air-permeable envelope 20 with the contacted while of the inner peripheral surface of syringe body portion 2, length is corresponding with the total length of slit 31 of removing bait portion 120.This air-permeable envelope 20 can be made of the material (as the film that poromeric material be made of) also higher than the ventilation property of the main material of slit 31.This air-permeable envelope 20 blocks above-mentioned OBL slit 31, and partly exposing from this OBL slit 31 to the part of outside becomes the breathable zones 21 that is made of a plurality of ventilation property unit areas.

In addition, can air-permeable envelope 20 not made tubular yet, but make OBL film, it is thickly blocked the outside attach configuration of the mode of each slit 31 from cylindrical body 3 according to liquid.

As above-mentioned poromeric material, can use with respect to cell suspending liquid 100 to be the resin permeability of non-perviousness (that is, can keep the close property of liquid).This resin permeability can be silicone resin for example, poly--4-methyl-1-pentene (P4M1P), polyisoprene, polyhutadiene, ethylene-vinyl acetate copolymer, new LDPE (film grade) and polystyrene etc.These poromeric materials have ventilation property preferably in plastic material, if but thickness increase then ventilation property reduce.Therefore, the wall thickness of air-permeable envelope 20 is preferably 200 μ m or following usually, preferred especially 100 μ m or following.

In addition, as poromeric material, in addition also can utilize porous film, the aperture of this porous film is set in below the prescribed value, so that can prevent cell suspending liquid 100 escape to outside in, can prevent that bacterium from invading to the medium pollution of cell suspending liquid 100.At this moment, the hydrophobic macromolecular material of the optimal seeking of raw and processed materials of porous film.The aperture of porous film preferably is located at 1 μ m or following, more preferably 0.4 μ m or following.As this hydrophobic material, can utilize polytetrafluoroethylene (PTFE), tetrafluoraoethylene-hexafluoropropylene copolymer, polyethylene terephthalate (PET) and polypropylene (PP), polyethylene (PE) etc.In addition, can also use the hydrophobicity polyvinylidene difluoride (PVDF).

Here, material as above-mentioned air-permeable envelope 20, more preferably use above-mentioned hydrophobic material as the porous film material etc., the silicone resin of perhaps above-mentioned resin permeability material, polyethylene or polystyrene, reason is, by using these materials, air-permeable envelope 20 can have ventilation property and two kinds of performances of cell non-adhesive.See the material that is the cell non-adhesive in the above-mentioned hydrophobic material more.

Injector type cell handling device 1 preferably carried out sterilising treatment in advance before preserving cell suspending liquid 100.When preserving cell, cell suspending liquid 100 is stored between the piston head 43 and bait portion 120 of air-permeable envelope 20 inboards of cylindrical body 3 inside of syringe body 2 (with reference to Fig. 1 (C)).Cell suspending liquid 100 can be following state: as putting into from bait portion 120 to syringe body portion 2 inside, afterwards by coming liquid thickly to keep 120 sealings of bait portion with lid 60 or resin; Perhaps, from rear end 12 sides of cylindrical body 3 cell suspending liquid 100 is put into afterwards earlier with 120 sealings of bait portion front end.

Though do not illustrate here, in injector type cell handling device 1, when having stored cell suspending liquid 100, also can store some gases.But, if becoming bubble, this gas sneaks into to nutrient solution, then might destroy cell, therefore preferably do not contain gas as far as possible.

1-2. the effect of injector type cell handling device 1

In the injector type cell handling device 1 of embodiment 1, cylindrical body 3 is by constituting for the material of non-perviousness with respect to cell suspending liquid 100, be formed with slit 31 on its side face 30, on this zone, be formed with breathable zones 21, its be with have pair cell suspension 100 for non-perviousness but the material that gas is had the character of perviousness form.Thus,, therefore can use above-mentioned various material, easily make by common operations such as injection forming because cylindrical body 3 needn't one is decided to be ventilation property.And, also guarantee easily as necessary rigidity of syringe and intensity.

In addition, this injector type cell handling device 1 is except as mentioned above syringe body portion 2 internal reservoir cell suspending liquids 100, and cell suspending liquid 100 is also keeping and can carry out gaseous interchange by breathable zones 21 and extraneous gas.Therefore, cell can absorb the necessary oxygen of existence from injector type cell handling device 1 outside, and can keep the pH of nutrient solution by carbonic acid gas, has therefore suppressed active reduction, can preserve well.

The bubble that exists in the cell suspending liquid 100 as contacting with cell, then may become the reason of destroying cell when carrying cell handling device.In the injector type cell handling device 1 of present embodiment 1, when breathable zones is used porous film, this bubble can be rejected to the outside with bubble antipollution simultaneously by breathable zones 21, so cell can be not destroyed, can be safely with its preservation.

And injector type cell handling device 1 can carry out gaseous interchange with the outside by being provided with breathable zones 21, but can not form from the mode that this breathable zones 21 is immersed into syringe body portion 2 inside according to bacterium.Therefore can antipollutionly take place, preserve cell well.The degree that prevents effect of this pollution as the material of air-permeable envelope 20, is suitably set its aperture when suitably setting its component thickness when for example using poromeric material or using porous film as previously mentioned, suitably adjusts thus.But this kind adjusting must be considered to have concurrently as the necessary ventilation property of cell handling device and carry out.

The above ventilation property effect that cell handling device of the present invention possessed, at the regenerative medicine scene, as outside hospital, gather cell and with its safe transport when possessing the functional body of clinical examination apparatus, be special requirement, the injector type cell handling device 1 of present embodiment 1 can be brought into play effective effect as the cell handling device that satisfies this requirement.

Therefore and injector type cell handling device 1 is made as the pattern of medical syringe, when using in regenerative medicine, the lid 60 of the front end of bait portion 120 is opened, and bait portion 120 is connected with pin, catheter in blood vessel or other conduits.Then, only push and cell can be injected in the organism with the pushing end 41 of finger with piston 40.Therefore, by with common injector operations operation much at one, do not need complicated operations, even if be not that the skilled personnel with high technology also can be injected into cell the affected part of organism, and can be easily and promptly implement regenerative medicine.

In addition, also can have following effect when using porous film to form breathable zones: by the pushing end 41 of piston 40 is gone into to 120 thrusters of bait portion, pair cell suspension 100 is exerted pressure, and can carry out the operation of bubble being removed from breathable zones 21 simply.

And, injector type cell handling device 1, when the cylindrical body 3 of syringe body portion 2 and in the air-permeable envelope 20 one or two are when being made of cell non-adhesive material, with the part of cell suspending liquid 100 contacted inwalls or on the whole cell do not adhere to.Thus, cell is saved with the state that suspends in nutrient solution, therefore when injector type cell handling device 1 is transplanted to cell in the organism, do not need to carry out operation with cell lift-off processing from cylindrical body 3 inwalls, can reduce the troublesome operation of this operation.In addition, because cell suspends, therefore only push piston 40 in nutrient solution, cell just can be transplanted in the organism from injector type cell handling device 1 inside swimmingly with nutrient solution.

Injector type cell handling device 1, by using above-mentioned cell non-adhesive material in syringe body portion 2, can reach fundamentally the effect of avoiding the problem of in the past following the cell lift-off processing, these problems promptly when physics is peeled off cell the damage problem of pair cell or make with medicament (strippers such as trypsinase, EDTA) when carrying out lift-off processing this medicament pair cell and transplant the organism of object dysgenic problem, must carry out the carrying out washing treatment of cell or utilize temperature variation to handle carrying out the loaded down with trivial details handling problem of cell lift-off processing etc. etc.

If use the injector type cell handling device 1 of present embodiment 1, then can carry out the Transplanted cells operation exactly on-the-spot very easy, rapid the reaching of regenerative medicine as mentioned above, even if therefore in the mechanism that does not possess so superb equipment, also can avoid the generation of problems such as polluting, implement regenerative medicine simultaneously well.Cell handling device of the present invention can satisfy well this in the operation of regenerative medicine desired condition.

As mentioned above, be injected in organism affected part or the blood vessel by the cell that will be kept at injector type cell handling device 1 inside, can be applied in the treatment to arthritis deformans, rheumatoid arthritis, pseudarthrosis, progressive muscular dystrophy, myocardial infarction, cerebral apoplexy, Parkinson's disease, Spinal injury, tendon injury, diabetes, hepatic insufficiency, digestion organs dysfunction, skin injury, leukemia, blood class disease etc.

In general, cell is cultivated with nutrient solution (perhaps substratum) and is preserved, utilize the injector type cell handling device 1 of present embodiment 1, when directly being injected in the organism, need to select that organism is had the nutrient solution that safety is formed with cell suspending liquid.

For example, when injecting chondrocyte or its precursor cell, when parkinsonian's brain injects neurocyte or its precursor cell or when the cardiac injects the myocardial cell, when the people injects cell etc. under the situation, preferably do not contain the substratum that organism such as bovine serum comes the synthetic medium of derived components or used the serum of patient own to the arthritis deformans patient.

(embodiment 2)

Figure 2 shows that sectional view as the structure of the injector type cell handling device 1 of the embodiment 2 of cell handling device one example of the present invention.Different with embodiment 1, in syringe body portion 2, on the perisporium of cylindrical body 3, communicating pores is not set, but on the front end face 1100 of this cylindrical body 3, forms breathable zones.This breathable zones, specifically, when forming syringe body portion 2, can be that only front end face 1100 parts are constituted, also can be formed by following mode by poromeric material: on front end face 1100, offer communicating pores, fusion adhesion or bonding air-permeable envelope cover this communicating pores, set thus to be the close property of liquid.

Above-mentioned cell non-adhesive material and above-mentioned air-permeable envelope material can use and the embodiment 1 listed material identical materials of enumerating.Usually be applied with as shown in Figure 2 at bait portion front end 120 places and cover 60, perhaps with resin with seal inside, liquid thickly keeps injector type cell handling device 1 inside thus.

The injector type cell handling device 1 of the embodiment 2 by having this spline structure, also can reach with much at one effect of embodiment 1 (gaseous interchange, bubble removed from cell suspending liquid 100 and can be easily and promptly carry out effects such as Transplanted cells).

In addition, the injector type cell handling device 1 of present embodiment 2 also has following higher effect: contact with cell suspending liquid 100 though promptly the inwall of syringe body portion 2 is most of, but because this cylindrical body 3 is made of cell non-adhesive material, therefore cell can not adhere to the inwall of syringe body portion 2, can preserve with the state that suspends in nutrient solution.

The injector type cell handling device 1 of present embodiment 2 is different with embodiment 1, on the cylindric inwall of syringe body portion 2 communicating pores is not set, therefore when the operation of removing bubble, with bait portion 120 up, from syringe body portion 2 inside that piston 40 is fashionable to 120 thrusters of bait portion, can easily bubble be concentrated near the front end face 1100, when leakage is suppressed at minimum, can easily this bubble be removed from front end face 1100, therefore preferred.Like this, can reach and prevent that bubble from sneaking into to cell suspending liquid 100, well cell is transplanted to the effect on biological side simultaneously.

Piston head 44 also can be made of poromeric material.Like this, cell suspending liquid 100 can carry out gaseous interchange with the outside more well, and is therefore preferred.

In addition, the also preferred cell non-adhesive material that uses constitutes piston head 44, makes great efforts to make cell not adhere to.

(embodiment 3)

Figure 3 shows that the sectional view of structure of injector type cell handling device 1 of the embodiment 3 of cell handling device of the present invention.With the difference of above-mentioned embodiment 1 and 2 be that it has following feature: syringe body portion 2 by constitute with the same cylindrical body of syringe body in the past, piston head 44 is made of poromeric material.Poromeric material can use the listed material of enumerating in above-mentioned embodiment 1 or 2.

Piston head 44 is by the chimeric syringe side front end that is fixed in the main body 42 of piston 40, so that liquid thickly is connected in syringe body portion 2 inwalls.The piston head 44 here only optionally sees through gas, so the solution of cell suspending liquid 100 can not escape to the outside.

Also preferred above-mentioned cell non-adhesive material formation syringe body portion 2 and the piston head 44 of using in the present embodiment 3.

The injector type cell handling device 1 of the embodiment 3 by having such formation also can reach and embodiment 2 effect much at one.

The injector type cell handling device 1 of present embodiment 3, has the piston head 44 of ventilation property by this, can prevent the generation of the pollution that bacterium etc. causes, the cell in the cell suspending liquid 100 can carry out gaseous interchange with the outside simultaneously, therefore can suppress to preserve cell well when cytoactive reduces.In addition, can also obtain following effect: promptly effectively the bubble that contains in the cell suspending liquid 100 is removed from piston head 44 and go to the outside, thereby prevent the generation of the cytoclasis that bubble causes.

When the injector type cell handling device 1 of use present embodiment 3 is removed the operation of bubble, opposite with embodiment 2, preferably bait portion 120 is operated downwards, its reason is the bubble in the cell suspending liquid 100 is concentrated near the air-permeable envelope of piston head 44, is easy to discharge.

In the present embodiment 3, provided the structure example that piston head 44 integral body all have ventilation property, but the present invention is not limited thereto, for example can also utilize elastomeric element same to make the main body of piston head earlier, at its interarea communicating pores is set, the mode that partly covers according to the perforation that will form thus is provided with breathable zones (for example air-permeable envelope), thereby constitutes piston head.

(embodiment 4)

Figure 4 shows that the outside drawing of structure of injector type cell handling device 1 of the embodiment 4 of cell handling device of the present invention.Present embodiment 4 is characterised in that, syringe body portion 2 is made of the non-adhesion material of above-mentioned cell, in syringe body portion 2, do not establish simultaneously breathable zones, and be provided with the lid 130 of filtering membrane at bait portion 120 front ends of syringe body portion 2, this filtering membrane with air-permeable envelope 131 as filtration unit.Material of air-permeable envelope (poromeric material) and cell non-adhesive material all can use the listed material of enumerating in the respective embodiments described above 1～3.

Have in the injector type cell handling device 1 of present embodiment 4 of this formation, by covering the air-permeable envelope 131 in 130, the inside and outside of injector type cell handling device 1 also can carry out gaseous interchange, therefore reached the effect almost same with the respective embodiments described above 1～3.

The bubble that removes of the injector type cell handling device 1 of embodiment 4 is operated also same with embodiment 3, to cover 130 earlier up, push piston 40, the bubble in the cell suspending liquid 100 is concentrated on around the bait portion 120 (and around air-permeable envelope 131) thus, it can be removed well.

The structure of the injector type cell handling device 1 of above-mentioned embodiment 2,3,4 is for to be provided with breathable zones at the place ahead or the rear of piston 40 slip directions.By the place ahead or the rear that is arranged on slip direction like this, be accompanied by the slide of piston 40, can easily bubble be driven out of from this breathable zones.

Here, in air-permeable envelope 20,131, front end face 1100 or the piston head 44 that in embodiment 1～4, lists, by using the porous film that constitutes by above-mentioned hydrophobic material, perhaps use resin permeability materials such as silicone resin, polyethylene or polystyrene, can have ventilation property and cell non-adhesive simultaneously, make cell better suspension, storage or carry out Transplanted cells swimmingly in cell suspending liquid 100.

(with respect to the variation and the performance comparison test of embodiment 1～4)

In above-mentioned embodiment 1～4, provided the example that constitutes syringe body portion 2 integral body by cell non-adhesive material, but not necessarily leave no choice but use this cell non-adhesive material to constitute syringe body portion 2 integral body among the present invention, only get final product with cell suspending liquid 100 contacted inwalls with cell non-adhesive material formation.In addition, even if use cell non-adhesive material partly to constitute inwall with the contacted syringe body of cell suspending liquid 100 portion 2, also can expect appropriate effect.But, in order to obtain effect of the present invention fully, the preferred cell non-adhesive material that still uses as far as possible constitutes integral body with the contacted syringe body of cell suspending liquid 100 portion 2 inwalls, perhaps uses cell non-adhesive material to constitute the integral body of syringe body portion 2.

The performance comparison test

Here for the relatively syringe and the performance of syringe of the present invention of prior art, serve as that cell cultures is carried out on the basis in each syringe with the pattern of following a～g, the survival rate of calculating cell.As the present invention's product, used the syringe of embodiment 2 and 4.

Fig. 5 is for showing the data of experimental result.

*****

A. the culture dish that uses suspension cell culture to use is cultivated

B. use the medical syringe of existing structure (polypropylene system), the front end of bait portion is covered, under the state of sealing, cultivate

C. use the medical syringe (front end face is provided with air-permeable envelope, syringe body portion is made by cell non-adhesive material) of embodiment 2, the front end of bait portion is covered, under the state of sealing, cultivate

D. use the medical syringe (front end face is provided with air-permeable envelope, syringe body portion is made by cell non-adhesive material) of embodiment 2, the lid that has filtering membrane (small-sized) of embodiment 4 is installed, under this state, cultivate at the front end of bait portion

E. use the medical syringe (front end face is provided with air-permeable envelope, syringe body portion is made by cell non-adhesive material) of embodiment 2, the lid that has filtering membrane (large-scale) of embodiment 4 is installed, under this state, cultivate at the front end of bait portion

*****

(area is 21cm in the cultivation of carrying out in culture dish in the suspension cell culture of a ²), as the cultivation in suspension system.

It is that 70 μ m, aperture are the film of 0.1 μ m that filtering membrane uses e-PTFE system, thickness.

Each size is as follows.

The air-permeable envelope area that has the lid of small-sized filtering membrane: about 43mm ²

The air-permeable envelope area that has the lid of large-scale filtering membrane: about 113mm ²

Syringe internal volume: 10mL

Ventilation property front end face area: about 70mm ²

*****

Here, the concrete evaluation method of cells survival rate is as described below.

Promptly, surface markers Leneage negative Scal+sKit+ (the following KSL: the suspension system cell) carry out purifying that will hemopoietic stem cell (HSC) concentrating and separating from mouse bone marrow cells comes out, 10000 of KSL cell count are suspended in the 2mL nutrient solution, the inner cultivation of each syringe 3 days, use the HSC number of surface markers and colony assay evaluation survival afterwards.

The kind of nutrient solution is as follows.

StemSpan substratum (Stemcell Technology)

The 50ng/mL mouse stem cells factor (mSCF; Peprotech)

20ng/mL people flt-3 part (Peprotech)

The 20ng/mL mouse platelets generates plain (mTPO; Stemcell Technology)

The microbiotic kind of adding: 0.05 μ g/mL Streptomycin sulphate

By experimental result shown in Figure 5 as can be known, at first in the b that has used existing medical syringe, cell inactivation is serious, and is relative therewith, obtained the highest performance as the d of the syringe of embodiment and the pattern of e.This result shows, in the syringe of embodiment, by air-permeable envelope that is arranged at syringe body portion front end face and the lid that is installed in the band filtering membrane of bait portion, can obtain higher gaseous interchange, can carry out good gaseous interchange with respect to the cell suspending liquid that is stored in syringe inside, its result has brought into play excellent cell retention.

So in the present invention, if make up the characteristic of the cell handling device of each embodiment mutually, then can expect to obtain very high cell retention.For example, in above-mentioned data example with embodiment 2 and 4 the combination examples, but the combination by carrying out breathable zones being set, making piston head have ventilation property as enforcement mode 3 in syringe body portion side as the enforcement mode 1 can have more large-area breathable zones.Thus, the cell in the cell suspending liquid can carry out gaseous interchange more well with the outside, reduces thereby suppress cell activity.In addition, the efficient of removing bubble from cell suspension also improves, and can not cause cytoclasis and preserves cell more safely.

Shown in c as can be known, even if when the lid of mounting strap filtering membrane not, by on the front end face of syringe body portion, air-permeable envelope being set, also can keep at least with cell cultures with the culture condition in the culture dish roughly the same or better cytoactive.On the other hand, consider that the cytoactive among the b descends,, can not obtain the such good cell retention of the present invention even only use existing medical syringe as can be known as cell handling device.

By above investigation as can be known, have simultaneously when Transplanted cells can be rapidly and easily with the performance and the high-caliber cell retention of Transplanted cells for the injector type cell handling device shown in the embodiment 1～4.

(embodiment 5)

Fig. 6 is the outside drawing of the structure of the cell handling device 200 of demonstration embodiment 5.

This cell handling device 200 is by having ventilation property and flexual material constitutes, and integral body is cylindric, and the surrounding wall portion 201 of its main body forms the snake shapes, can stretch when keeping drum thus.The leading section of main body surrounding wall portion 201 is formed with bait portion 203, chimericly when not using covers 60, and liquid thickly keeps main body surrounding wall portion 201 inside thus.Cell suspending liquid 100 (not shown) is stored in the inside of cell handling device 200.

Material as constituting cell handling device 200 can use the poromeric material that lists in embodiment 1～4.But, in the present embodiment 5,, therefore, considering that the shape with cell handling device remains under the situation of drum because this poromeric material has almost constituted the integral body of cell handling device, the preferred use has suitable inflexible material.In addition, when using porous film as the material of cell handling device 200, the transparency of cell handling device descends, and can't confirm cell suspending liquid 100 wherein sometimes.At this moment, can use the part (rearward end 202 of main body surrounding wall portion 201) of resin permeability material formation cell handling device 200, guarantee the transparency of cell handling device, make it possible to confirm to be stored in inner cell suspending liquid 100.Cell handling device 300 to aftermentioned embodiment 6 also can use this way.

The cell handling device 200 of the embodiment 5 by having said structure, because these cell handling device 200 integral body are all made by above-mentioned poromeric material, therefore can prevent that liquid thickly is stored in the generation that inner cell suspending liquid 100 is polluted by bacterium etc., and the broad area that can spread all over cell handling device 200 inwall integral body absorbs the required oxygen of cells survival.Therefore, compare for the cell handling device of ventilation property, can significantly obtain good gaseous interchange performance with only a part of.Therefore particularly, when preserving, also can easily keep original container form because cell handling device 200 forms the snake shape, and, can guarantee to enlarge the surface-area of the cell handling device relevant with the gaseous interchange of cell.

And cell handling device 200 is compared with the formation of the respective embodiments described above 1～4, also has advantage simple in structure, easy to manufacture.And, because cell handling device 200 do not need piston, in therefore also needn't worrying to preserve owing to leakage (sealing leakage) takes place the gap between syringe body portion and the piston.

Also performance to some extent during the use of the excellent properties of this cell handling device 200 in regenerative medicine.Promptly, to cover 60 during use opens, pin or conduit are installed at bait portion 203 places, the prescribed position of corresponding organism inside, only by manually or the rearward end 202 of anchor clamps pushing main body surrounding wall portion 201, just can avoid under the situation that pollution takes place easily and promptly the cell of inside is being injected into biological side.In addition, when the part of device is used porous film, also can carry out the operation that the bubble in the cell suspending liquid 100 is removed effectively by pushing rearward end 202.

(embodiment 6)

Figure 7 shows that the outside drawing of the structure of the cell handling device 300 that shows embodiment 6.

This cell handling device 300 by with above-mentioned cell handling device 200 same have ventilation property and flexual material constitutes, its main body surrounding wall portion 301 forms elongated bag (pipe) shape.The leading section of main body surrounding wall portion 301 is formed with bait portion 303, chimericly in this bait portion 303 when not using covers 60, and liquid thickly keeps inner thus.And, in main body surrounding wall portion 301, insert logical flat cyclic operation ring 304 from rearward end 302 sides.Cell suspending liquid 100 (not shown) liquid thickly is stored in the inside of cell handling device 300.

The cell handling device 300 of the embodiment 6 by having this structure also can reach and embodiment 5 effect much at one, but because structural difference, particularly in cell handling device 300, owing to do not need cell handling device 200 as embodiment 5, be processed into snake shape etc., therefore also have and make the advantage that becomes and be more prone to.

By this cell handling device 300, when in regenerative medicine, using, at first will cover 60 and open, pin or conduit are installed in bait portion 303, keep the rearward end 302 of main body surrounding wall portion 301.Then, with pointing move operation ring 304 forwards.Only by carrying out this simple operation, be subjected to operating the pressure of ring 304, main body surrounding wall portion 301 is urged, and just can cell suspending liquid 100 be discharged from bait portion 303.Thus, almost same with embodiment 5, can reach when avoid to take place polluting, easily and promptly inner cell is injected into the effect on biological side.

In the above-mentioned embodiment 5 and 6, cell handling device integral body is for the snake shape or have flexual elongate pocket shape, but cell handling device of the present invention is not limited thereto shape, also can be for example cylindric, spherical, rectangular-shaped, as long as the material of cell handling device, make getting final product at least by the material that lists as the constituent material of above-mentioned cell handling device 200,300.In such cases,, cell can be migrated to organism well when then using on this cell handling device, realize regenerative medicine rapidly if bait portion is set, therefore preferred.

In the embodiment 5 and 6, material as cell handling device 200,300, same with the listed air-permeable envelope of enumerating 20 and 131 in the embodiment 1～5, front end face 1100, piston head 44, cell handling device 200, by using above-mentioned porous film or the resin permeability materials such as silicone resin, polyethylene or polystyrene that constitute by hydrophobic material, make cell handling device 300 have the cell non-adhesive, then cell or cell mass can suspend more well, store in cell suspending liquid 100 or carry out Transplanted cells swimmingly.

In the above-mentioned embodiment 5 and 6, because reasons such as the shelf time of cell suspending liquid 100 is extremely short, when not requiring ventilation property so, so that cell or cell mass are suspended in the effect of preserving in the nutrient solution is main purpose, in the material of cell handling device 200,300, can use the listed cell non-adhesive material of enumerating of material as the syringe body portion 2 of embodiment 1.

Snake shape, the piped cell handling device 200,300 that lists in above-mentioned embodiment 5 and 6 can also be utilized as the capsule 50 of syringe body portion 2 inside that are accommodated in embodiment 7,8 described later.

(embodiment 7)

Figure 8 shows that the sectional view of the structure of the injector type cell handling device 1 that shows embodiment 7.

Possess the syringe body portion 2 and the piston 40 that are constituted by cylindrical body 3 and capsule 50 in the structure of this injector type cell handling device 1.

Cylindrical body 3 and piston 40 can use the material that is generally used for syringe to make.Cylindrical body 3 forms tubular roughly, and the front end side is formed with the discoid front end face 110 that central part has perforated portion 11.

Capsule 50 forms according to the mode that two ends have bait portion 51 and rearward end 52, is the liquid memory of the volume pliability cylindrical shell that can dwindle.The rearward end 52 and the piston crown 43 of this capsule 50 dispose in opposite directions, by pushing its integral body, become the reducible pushing of volume portion, and by pushing with piston head 43, the internal reservoir thing is pushed out from bait portion 51.In addition, take in bait portion 51, make its perforated portion 11 be exposed to the outside more outerly, when being pressed to the place ahead, storing thing and be difficult to residue in the capsule 50 with piston crown 43 than above-mentioned front end face 110 along the inner peripheral surface of cylindrical body 3.Bait portion 51 is preferably by forming than the harder material forming in position that stores cell, so as can cover lid, conduits such as pin or conduit are installed.

The injector type cell handling device 1 of present embodiment 7 has cell suspending liquid 100 and thickly is stored in structure in this capsule 50 by liquid.Here in the present embodiment 7, this capsule 50 has principal character.Be that capsule 50 is made of the material that has close property of liquid and ventilation property simultaneously, the cell that is stored in thus in the inner cell suspending liquid 100 can not flow out to bait portion 51 part in addition, and can carry out gaseous interchange with the outside of capsule 50.

As this poromeric material, can use the listed poromeric material of enumerating in above-mentioned embodiment 1～6.

In addition, capsule 50 desired ventilation properties are different and different according to surface-area, cell loading level, cell category and the preservation condition etc. of cell handling device, but must have the sufficient amount that the cell that is filled in cell handling device is used to survive.Ventilation property as long as have in capsule 50 upper inside wall with dividing, but in order fully to obtain ventilative efficient, the integral body of preferred capsule 50 inner wall section or the integral body of capsule 50 are made of poromeric material.

In the time of in the injector type cell handling device 1 that cell suspending liquid 100 is filled to present embodiment 7, can place empty capsule 50 in the inside of cylindrical body 3 earlier, from bait portion 51 front ends cell suspending liquid 100 is stored in the inside of capsule 50 then; Also tytosis suspension 100 in capsule 50 earlier places it in the inside of cylindrical body 3 then.Like this, use in order to make capsule 50 to take out from cylindrical body 3, the size of the perforated portion 11 of front end face 110 must following setting: be set to be equipped with and cover 60 bait portion 51 and can insert logical size; Perhaps when capsule 50 being installed on the cylindrical body 3 back mounting cover 60 or when the perforated portion 11 that cover 60 bait portion 51 insertion front end faces 110 will be installed, being set at and being deformed into insertable size and shape.

When not using (during preservation), at the chimeric lid 60 of the front end of bait portion 51, liquid thickly keeps capsule 50 inside thus.

With in bait portion 51 well mounting cover 60 be purpose, preferably in capsule 50, constitute bait portion 51 with having to a certain degree the inflexible material at least.

Piston 40 is all same with embodiment 1, but has following feature, promptly on the elastomeric element that constitutes piston head 43, is formed with a plurality of holes 431 along the syringe direction of principal axis.As stream, make great efforts to make the cell suspending liquid 100 of capsule 50 inside to carry out gaseous interchange in this hole 431 with capsule 50 outsides (from the outside outside of piston head 43).

This injector type cell handling device 1 uses through after the sterilising treatment basically.

First being characterized as of injector type cell handling device 1 with embodiment 7 of this formation, capsule 50 is preventing to have ventilation property when the bacterium intrusion from waiting the pollution that causes, the cell that is contained in the cell suspending liquid 100 of capsule 50 inside can be situated between by capsule 50, carries out gaseous interchange from the hole 431 of piston head 43 and gap and the outside between bait portion 51 and the perforated portion 11.Therefore, the cell in the cell suspending liquid 100 can absorb the necessary oxygen of existence from the outside of capsule 50, thereby can activity not reduce and be preserved well in capsule 50.

In the present embodiment 7, hole 431 with piston head 43, with the perforated portion 11 of front end face 110 as stream, can carry out gaseous interchange, as long as but can provide gas from installing outside pair cell, certainly not only be defined in hole 431, perforated portion 11, can form breathable zones at other any position.

Material as the capsule 50 of present embodiment 7 and embodiment described later 8, can be same with the listed air-permeable envelope of enumerating 20 and 131 in the embodiment 1～6, front end face 1100, piston head 44, cell handling device 200 and 300, use the above-mentioned porous film that constitutes by hydrophobic material, perhaps use resin permeability materials such as silicone resin, polyethylene or polystyrene, capsule 50 also has the cell non-adhesive thus, can make cell suspend more well in cell suspending liquid 100 and stores.Like this, do not need as in the past, when cell is taken out, to make with medicament (strippers such as trypsinase, EDTA), handle the troublesome operations that cell is peeled off by temperature variation from cell handling device, can prevent the physics and the chemical damage of cell, improve operation efficiency greatly.In addition,, therefore the disposal of regenerative medicine rapidly can be carried out, burden can also be alleviated the patient who accepts Transplanted cells owing to do not need troublesome operation such as this lift-off processing, cell washing processing.

Second of the injector type cell handling device 1 of embodiment 7 is characterized as, owing to be made into the medical syringe type, therefore when in regenerative medicine, using, same with the injector type cell handling device 1 of embodiment 1～4, can be rapidly and easily carry out the operation of Transplanted cells.

When capsule 50 is when being made of porous film, be set in the size of afore mentioned rules by the aperture with film this moment in the injector type cell handling device 1 of present embodiment 7, also be difficult for taking place leakage even if capsule 50 exerted pressure.Therefore, the cell suspending liquid 100 of preciousness spills loss in the time of can preventing to remove bubble.This effect is effective especially when requiring to guarantee cell concentration.

When constituting capsule 50 by porous material, capsule 50 can bleach under the different situations, becomes and can not confirm the inside of capsule 50, therefore should be noted that.The visibility of capsule 50, especially in the residual content of confirming cell suspending liquid 100, to have when pollution-free be essential, therefore in this case, can carry out operations such as adjustment apertures rate, aperture.

The injector type cell handling device 1 of present embodiment 7, cylindrical body 3 does not contact with cell suspending liquid 100 with piston 40 at least, therefore can utilize again.

Above-mentioned example is for using poromeric material or having ventilation property and the material of cell non-adhesive formation capsule 50, wait when shelf time of cell is short, when the requirement of ventilation property is not so important, also can uses constituent material, constitute capsule 50 as the cell non-adhesive material of main purpose with above-mentioned cell non-adhesive as cylindrical body 3.

(embodiment 8)

Figure 9 shows that the sectional view of the structure of the injector type cell handling device 1 that shows embodiment 8.

The difference of present embodiment 8 and above-mentioned embodiment 7 be following some: be not provided with hole 431 on piston head 43, in the inside of cylindrical body 3, the capsule leading section is 53 sealed, in the inside of cylindrical body 3, with capsule leading section 53 capsule is set in opposite directions and destroys mechanism 13.

Capsule destroys mechanism 13 and specifically can be made of sharp steel edge or needle-like member.Among Fig. 9 example destroy mechanism 13 and be provided with the example of steel edge as capsule, but when needing to consider waste treatment, this capsule destroys mechanism 13 can be by for example constituting with syringe body portion same resin component.

Utilization has the injector type cell handling device 1 of the embodiment 8 of this structure, also can reach injector type cell handling device 1 effect much at one with embodiment 7 when cell is preserved.

As shown in Figure 9, the injector type cell handling device 1 of present embodiment 8, because the peristome 121 of bait portion 120 that will be located at front end face 110 is as stream, and the leading section 53 of capsule 50 exposes externally in the air, therefore compare with embodiment 7, have the effect that to carry out gaseous interchange more well and remove bubble.

(with respect to the variation of cell handling device)

Example shown in the shape of the cell handling device of explanation is not limited to certainly in above-mentioned embodiment 1～8 can also be other shape.That is, but so long as at least inside liquid thickly store the cell handling device of cell, at least with the contacted cell handling device inwall of above-mentioned cell in a part or mostly be to constitute by cell non-adhesive material or poromeric material to get final product.

For example, cell handling device of the present invention can be columned face shaping.At this moment, in this cell handling device, preserve cell earlier, can pin or conduit be installed in the side of cell handling device during use, make the volume-diminished of cell handling device simultaneously, cell is discharged.

2. about tissue regeneration composition of the present invention

Figure 10 shows that the partial enlarged drawing of tissue regeneration of the present invention with composition.This tissue regeneration is made of cell cultures particle 1000 and the mobile medium 2000 (for example nutrient solution) that contains it with composition.

This tissue regeneration is as described below with composition, can replace in the past being used with composition by support with fairly large structure and the tissue regeneration that cell constituted that distributes on this support, the tissue regeneration of using as regenerative medicine has the suitableeest performance with composition.

As shown in the drawing like that, cell cultures particle 1000 has following formation: promptly on a plurality of surfaces that are scattered in scaffold particle 1001 in the mobile medium 2000, that formed by bioabsorbable material, be stained with a plurality of cells 1002.The adhesion of cell 1002 is several to be changed according to each scaffold particle 1001.

The scope that scaffold particle 1001 preferred diameters are 10 μ m～2000 μ m is preferably the scope of 50 μ m～500 μ m especially.By setting particle diameter for this reason, cell cultures particle 1000 can disperse in mobile medium 2000 well, thereby all has flowability as tissue regeneration with composition in its entirety.Therefore, be kept at composition in the cell handling device of above-mentioned embodiment 1～8, then can be taken out to the outside well from this cell handling device inside by bait portion etc. if will contain the tissue regeneration of cell cultures particle 1000.In addition, cell cultures particle 1000 is a purpose with the adhered area of guaranteeing cell, is preferably to have the porous plastid that cell immerses the pore in required enough apertures.

As the above-mentioned bioabsorbable material that constitutes scaffold particle 1001, can use material known, be selected from aliphatic polyester as using, poly(lactic acid), polyglycolic acid, lactic acid-ethanol copolymer, lactic acid-caprolactam copolymer, oxyacetic acid-carbonate copolymer, poly-to the dioxy cyclohexanone, chitosan, cross-linked-hyaluronic acid, alginic acid, osso-albumin, laminine, fibronectin, external connection albumen, polylysine, scleroproein, calcium phosphate, lime carbonate, polybutylcyanoacrylate, polyglutamic acid, polyhydroxybutyrate, polymalic acid, poly-acid anhydrides, poe, chitin, starch, Fibrinogen, in hydroxylapatite and the gelatin a kind or multiple.These materials can use separately respectively also can exercise usefulness with multiple being combined into.

As the Production Example of this scaffold particle 1001, can solvent evaporation be removed make afterwards by will in solvent, being dissolved with solution spray, the corpusculed of above-mentioned bioabsorbable material; Can also make its emulsification in the dispersion solvent by material solution is dispersed in, then the method for solvent evaporation be made; Perhaps with the emulsifying raw material of above-mentioned bioabsorbable material, by its polymeric letex polymerization is made.

As the method for scaffold particle 1001 being made porous matter, can list bioabsorbable material solution emulsification, freeze-drying, follow freeze-drying with solvent evaporation.In addition, the mixture that is used in powder such as the salt that is mixed with in the bioabsorbable material as pore-forming material, sugar is in addition made particle, then the pore-forming material dissolving is removed, and thus particle is become the method for porous matter.

On the other hand, in cell 1002, can use above-mentioned various cell category according to therapeutic purpose.This cell category is not particularly limited, can list embryo's property stem cell (embryonic stem cells:ES cell) particularly, embryo's sexual reproductive cell (embryonic germ cells:EG cell), adult stem cell (adult stem cells:AS cell, be also referred to as adult stem), mescenchymal stem cell, neural stem cell, the blood vessel endothelium stem cell, hemopoietic stem cell, stem cells such as liver stem cells can also list osteocyte in addition, the chondrocyte, the myocyte, the myocardial cell, neurocyte, tendon cell, adipocyte, pancreatic cell, liver cell, nephrocyte, hair follicle cell, cell or its precursor cell that hemocyte etc. have broken up.Like this, can use with each differentiation degree headed by embryo's property stem cell different stem cell and the cell that is divided into each tissue.Wherein, when using adherent cell, because the support of propagation and differentiation phase can be provided, therefore effectively.Here, so-called adherent cell comprises the nearly all cell the hemocyte in hemopoietic stem cell.

In addition, can also use utilize the genetic engineering method will be here the listed gene alteration cell of enumerating any one change of various cells and forming.

And,, except general nutrient solution, also can use nutrient solution medium in addition, for example physiological saline, phosphoric acid buffer etc. as above-mentioned mobile medium 2000.But the nutrient solution and these media that use in the cell cultures must be replaced in such cases.

When making above-mentioned cell cultures particle 1000, at first can in nutrient solution (mobile medium 2000), above-mentioned cell 1002 be inoculated on the scaffold particle 1001, in this nutrient solution, cultivate.Thus, cell 1002 adheres to the surface of scaffold particle 1001 naturally, has constituted cell cultures particle 1000.Cell cultures can be carried out in accordance with known methods.

Example as the working order of cell cultures, at first from organism with cellular segregation and refining, the select target cell, as required add growth factor thereafter, make its propagation or to the target cell induction, in nutrient solution, add cell cultures particle 1000 simultaneously, and slowly stir and cultivate.At this moment, also can in the cell cultures particle, contain the necessary compositions of cell proliferation such as growth factor.

Here, Shi Ji cell cultures is preferably carried out in incubator.The tissue regeneration composition of gained is accommodated in the cell handling device listed cell handling devices of enumerating such as (preferred) embodiments of the present invention 1～8 of regulation, preserves until being used for the treatment of under the condition that is suitable for preserving.Preserve and preferably to carry out at low temperatures, also can under normal temperature or heating, carry out being difficult to cause under the condition of cell inactivation.

By this method, cell 1002 adheres in the lip-deep hole of porous matter of scaffold particle 1001, has constituted cell cultures particle 1000, culturing cell 1002 well in cell culture medium or induction substratum.

In order further to improve the adhesivity of 1002 pairs of scaffold particles 1001 of cell, preferably known method (physical treatments such as ozonize, UV processing, plasma treatment are passed through on scaffold particle 1001 surfaces, chemical treatments such as vitriolization, salt acid treatment, coating are handled etc.) processing of implementing to improve cell adhesion.As coating material, can list the formation that covers with laminine, fibronectin, external connection albumen, polylysine, scleroproein (comprise and solidify processing in advance), Fibrinogen and gelatin or osso-albumin.Physiologically active substances such as various multiplicaiton factor, hormone can also be fixed, be adsorbed to particle itself.Constitute by this, scaffold particle 1001 physical strengths own improve, and the adhesivity of cell 1002 also improves, and curative effect also may improve simultaneously.

(effect of particle shape tissue regeneration composition)

By having the tissue regeneration composition of above formation, cell 1002 can be in the surface of scaffold particle 1001 propagation or to the target cell induction, during this, cell 1002 becomes fine cell cultures particle 1000 with scaffold particle 1001, be the state of suspension in nutrient solution, integral body has flowability.And when the use of regenerative medicine, cell cultures particle 1000 can directly be injected into and carry out Transplanted cells in the organism.

Because scaffold particle 1001 is formed by bioabsorbable material, if therefore cell cultures particle 1000 is kept in the injector type cell handling device of explanation in the above-mentioned embodiment 1～8, then in use, this cell cultures particle 1000 can carry out Transplanted cells to organism from bait portion etc. swimmingly with mobile medium 2000.

As the method for carrying out Transplanted cells, have at the affected part of heeling-in support not and carry out method that cell injects, the support that is imbedded at affected part is in advance carried out method that cell injects etc. repeatedly by this injector type cell handling device.Back one method is effective under the situation that slowly makes the affected part refective regeneration (regenerating bone or cartilage, breast regrowth etc.).

Promptly, in general, when in container, preserving cell, cell easily adheres to the inwall of container in the preservation process, therefore be difficult to cell is taken out to outside the container, if but when being kept in the said syringe type cellular processor tool, then cell does not adhere to inwall in the preservation process, cell cultures particle 1000 keeps the state that suspends in mobile medium 2000.Therefore, connect pin, catheter in blood vessel,, just cell cultures particle 1000 can be injected in the organism by pushing piston etc. in the bait portion of cellular processor tool.

In said syringe type cellular processor tool, culturing cell on scaffold particle 1001 can utilize injector operations to be injected in the organism from the cellular processor tool by the cell cultures particle 1000 that cultivation forms.

Owing to scaffold particle 1001 disappears the therefore operation that does not need to remove support after transplanting through being absorbed by organism behind the certain hour in being injected into organism.Therefore, patient's burden is significantly reduced, particularly also can alleviate burden patients such as the relatively poor child of muscle power, old men.

If emboliform tissue regeneration is kept in the above-mentioned embodiment 1～8 in the listed cell handling device of enumerating with composition, then can prevent the cytoclasis that cell causes to adhesion, cell inactivation, the bubble of cell handling device inwall, preserve cell simultaneously well.

Be applicable to the example of regenerative medicine with composition as tissue regeneration of the present invention, by being injected into the affected part of organism, can be used in the treatment to arthritis deformans, rheumatoid arthritis, pseudarthrosis, periodontopathy, progressive muscular dystrophy, myocardial infarction, cerebrovascular disorder, Parkinson's disease, Spinal injury, tendon injury, diabetes, hepatic insufficiency, digestion organs dysfunction, skin injury, leukemia, blood class disease etc.

More particularly, inject chondrocyte or its precursor cell, inject osteocyte or its precursor cell, inject neurocyte or its precursor cell, inject myocardial cell or its precursor cell etc. to the arthritis deformans patient to the cardiac to parkinsonian's brain to the periodontopathy patient.

In the above explanation, illustration utilized the example of regenerative medicine as cell category with cell, but, can also use regenerative medicine treatment in addition cell or other cells as being used for cell handling device of the present invention and tissue regeneration cell with composition.

Cell handling device of the present invention and tissue regeneration composition, be injected in organism affected part or the blood vessel by the cell that will store, can be applicable in the treatment to arthritis deformans, rheumatoid arthritis, pseudarthrosis, progressive muscular dystrophy, myocardial infarction, cerebral apoplexy, Parkinson's disease, Spinal injury, tendon injury, diabetes, hepatic insufficiency, digestion organs dysfunction, skin injury, leukemia, blood class disease, hair regeneration etc.

Claims (41)

1. injector type cell handling device, wherein, it possesses and can thickly store the main part of the mobile process object thing that contains cell and insert the interior piston of main part slidably by liquid, this piston is applied in and is used for the cell that is stored in main part inside is transplanted to biological intravital thrust pressure, described injector type cell handling device has following formation: by destroying the sealed state of the main part that liquid thickly seals, the volume in the fluid storage space of main part is changed, make the process object logistics go into main part or flow out thus from main part; Breathable zones in the gas permeation that is provided with the cells survival institute necessary amounts that is used for making described process object thing at least a portion of described main part and/or piston, at least a portion zone at the position that contacts with described process object thing under the close state of liquid.

2. cell handling device as claimed in claim 1 is characterized in that, the ventilation property of described breathable zones is counted more than or equal to 1mL/24hratm to be converted into total oxygen permeating amount.

3. cell handling device as claimed in claim 1 or 2 is characterized in that described breathable zones is formed by resin permeability or porous film.

4. tissue regeneration method, it comprises second step that cell is stored in the first step in the container and the cell of preserving is transplanted to organism, it is characterized in that, in the described first step and/or second step, use each described cell handling device of claim 1～3.

5. cell handling device as claimed in claim 1, it is characterized in that, described main part is by the storage cell and along with the slip of piston bag-like container that can be out of shape arbitrarily and the urceolus portion of taking in this container constitute, and at least a portion of described bag-like container is described breathable zones.

6. cell handling device as claimed in claim 5 is characterized in that, described bag-like container can break away from from described outer cylinder body, can carry out the processing of cell under disengaged position.

7. cell handling device as claimed in claim 1 is characterized in that, described bag-like container possesses discharge portion of discharging cell and the pushing portion that this packed container is applied thrust pressure.

8. cell handling device as claimed in claim 7 is characterized in that, described bag-like container is except described discharge portion, and all the other all are made of the flexible material that can dwindle by pushing.

9. cell handling device as claimed in claim 8 is characterized in that described bag-like container possesses the part that forms the snake shape, and by the thrust pressure that described pushing portion applies, this snake shape partly dwindles.

10. cell handling device as claimed in claim 8 is characterized in that, described bag-like container is a tubulose.

11. cell handling device as claimed in claim 5, it is characterized in that, on the part beyond the bag-like container of described cell handling device, has described breathable zones, so that under described bag-like container is accommodated in state in the described urceolus portion, can carry out gaseous interchange in this bag-like container and between the treatment unit outside.

12. cell handling device as claimed in claim 5 is characterized in that, it possesses destruction mechanism, and this mechanism destroys described bag-like container under described bag-like container is accommodated in state in the described urceolus portion.

13. cell handling device as claimed in claim 5, it is characterized in that, in the described main part, be provided with the discharge portion that described cell or tissue regeneration are discharged with composition in the place ahead of piston slip direction, this discharge portion is according to forming with the mode that pin, catheter in blood vessel or other conduits are connected.

14., it is characterized in that the ventilation property of described breathable zones is counted more than or equal to 1mL/24hratm to be converted into total oxygen permeating amount as each described cell handling device of claim 5～13.

15., it is characterized in that described breathable zones is formed by resin permeability or porous film as each described cell handling device of claim 5～13.

16. tissue regeneration method, it comprises second step that cell is stored in the first step in the container and the cell of preserving is transplanted to organism, it is characterized in that, use each described cell handling device of claim 5～15 in the described the first step and/or in second step.

17. cell handling device as claimed in claim 1 is characterized in that, described breathable zones is arranged on a plurality of positions of described main part independently, and each ventilation property unit area extends setting along the slip direction of described piston.

18. cell handling device as claimed in claim 17 is characterized in that, described ventilation property unit area is formed by the higher material of ventilation property than the material of main part of described main part.

19. cell handling device as claimed in claim 1 is characterized in that, described breathable zones is positioned at along the position of described piston slip direction.

20. cell handling device as claimed in claim 19 is characterized in that, described breathable zones is formed at the leading section of described piston.

21. cell handling device as claimed in claim 19 is characterized in that, described breathable zones be positioned at described main part the discharge cell discharge portion near.

22. cell handling device as claimed in claim 21 is characterized in that, described discharge portion be formed at when pushing described piston fully and the contacted front end face of this piston or its near, described breathable zones is formed at described front end face.

23. cell handling device as claimed in claim 21 is characterized in that, described breathable zones is arranged in the packaged unit of blocking described discharge portion.

24., it is characterized in that the ventilation property of described breathable zones is counted more than or equal to 1mL/24hratm to be converted into total oxygen permeating amount as each described cell handling device of claim 17～23.

25., it is characterized in that described breathable zones is formed by resin permeability or porous film as each described cell handling device of claim 17～23.

26. tissue regeneration composition, it is characterized in that, it constitutes: the cytoskeleton particle with mobile medium and particle shape of the support that becomes the cell that suspends in this flowability medium, described cytoskeleton particle is made of bioabsorbable material, and this cytoskeleton particle has cell adhesion simultaneously.

27. tissue regeneration composition as claimed in claim 26 is characterized in that, the diameter of described scaffold particle is the scope of 10 μ m～2000 μ m.

28. as claim 26 or 27 described tissue regeneration compositions, it is characterized in that described scaffold particle is a porous matter.

29., it is characterized in that described scaffold particle has been implemented the processing that improves cell adhesion as claim 26 or 27 described tissue regeneration compositions.

30., it is characterized in that described cell is to be selected from the cell that needs when cell proliferation among the adherent cell of the support group as claim 26 or 27 described tissue regeneration compositions.

31. cell handling device that has tissue regeneration with composition, it is characterized in that, can thickly store the mobile process object thing that contains cell by liquid, at least the part at position of contacting with described cell is provided with the breathable zones of the gas permeation that is used to make cells survival institute necessary amounts, as described process object thing, this tissue regeneration contains mobile medium with composition and becomes the support of the cell that suspends and the cytoskeleton particle of the particle shape that is made of bioabsorbable material in described mobile medium the regeneration of this cell handling device stored tissue with composition.

32. the cell handling device that has tissue regeneration with composition as claimed in claim 31 is characterized in that the whole of the part of the storage cell of described at least cell handling device are described breathable zones.

33. the cell handling device that has tissue regeneration with composition as claimed in claim 31, it is characterized in that, described cell handling device possesses the volume that the spatial volume that stores cell is changed and changes mechanism, discharges or flow into cell along with changed the volume-variation that mechanism causes by described volume.

34. the cell handling device that has tissue regeneration with composition as claimed in claim 31, it is the injector type cell handling device that has the main part that stores cell and be inserted into the piston in the main part slidably, this piston is applied in and is used for being transplanted to the thrust pressure of organism with being stored in inner cell, it is characterized in that at least a portion of described main part and/or piston is described breathable zones.

35., it is characterized in that with comparing with described breathable zones with the contacted position of the cell of described main part, cell is easier to adhere on the described cytoskeleton particle as claim 31 or the 34 described cell handling devices that have tissue regeneration with composition.

36. the cell handling device that has tissue regeneration with composition as claimed in claim 35, it is characterized in that, described main part is provided with in the place ahead of the slip direction of piston discharges the discharge portion of described cell or tissue regeneration with composition, and this discharge portion is connected with pin, catheter in blood vessel or other conduits.

37. the cell handling device that has tissue regeneration with composition as claimed in claim 35 is characterized in that described main part and/or piston are and the contacted position of described cell that a part at least wherein is made of cell non-adhesive material.

38. the cell handling device that has tissue regeneration with composition as claimed in claim 31 is characterized in that the ventilation property of described breathable zones is counted more than or equal to 1mL/24hratm or more than or equal to 10mL/24hratm to be converted into total oxygen permeating amount.

39. the cell handling device that has tissue regeneration with composition as claimed in claim 31 is characterized in that described breathable zones is formed by resin permeability or porous film.

40. the cell handling device that has tissue regeneration with composition as claimed in claim 37 is characterized in that described cell non-adhesive material is any in hydrophilic material, hydrophobic material or the material that has negative charge.

41. tissue regeneration method, it comprises second step that cell is stored in the first step in the container and the cell of preserving is transplanted to organism, it is characterized in that, in the described first step and/or second step, use each described cell handling device that has tissue regeneration with composition of claim 31～40.

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