CN103614296A - Double-chamber three-dimensional pouring bioreactor system - Google Patents
Double-chamber three-dimensional pouring bioreactor system Download PDFInfo
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Abstract
The invention discloses a double-chamber three-dimensional pouring bioreactor system which comprises a bioreactor, wherein the bioreactor is internally provided with a three-dimensional bracket and is connected with a pouring device. The bioreactor comprises a first chamber filled with common nutritional nutrient liquor and a second chamber filled with inductive culture factors. A dialysis membrane assembly through which components having molecular weight being less than the molecular weight of inductive culture factors in the inductive culture factors can pass is arranged between the first chamber and the second chamber. The three-dimensional bracket is located in the second chamber. According to the double-chamber structural design of the bioreactor disclosed by the invention, the common nutritional nutrient liquor and the inductive culture factors in the two chambers are independently exchanged, and the inductive culture factors can be preserved while the common nutritional nutrient liquor is changed, so that the use level of the inductive culture factor is greatly reduced, the effective utilization ratio of the inductive culture factors is improved, and the cost is lowered.
Description
Technical field
The invention belongs to biotechnology apparatus field, especially relate to the two-chamber dimensional perfusion bioreactor system of cell Growth and Differentiation in three-dimensional rack.
Background technology
At present, in life science field and medical field, cell cultures mode has multiple, by the dimension classification of cultivating, substantially can be classified as two dimension and dimensional culture mode.In two dimension training method, as the culturing bottle of glass or plastics, culture dish, need to provide a plane for cell attachment growth, by providing cell better growing environment to the modification of the classification of planar materials and contact surface, equally by inverted microscope and the simple microscope growth conditions of observation of cell very easily.But normal cell is all to grow in three-dimensional environment in human body and animal body, some cell cannot adhere to any planar growth, therefore for cell, provide a kind of environment of three-dimensional to cultivate more to meet in vitro the normal physiological state of cell, contribute to the corresponding process of growth in better analogue body.
Dimensional culture is having various ways in technical field at present, as: (1) cultivates on the basis of plane in two dimension, side adds with one deck collagen matrices composition in the plane, cell is grown in the three-dimensional environment of matrix, also can make three-dimensional rack by artificial material, in conjunction with perfusion system, cultivate; (2) swing of revolving oar and the rotation by different shape provides power, cell suspension is grown in the three-dimensional environment of cultivating liquid, also have by some artificial materials single or a plurality of cell micro-encapsulations, for cell provides attachment surface, contact with intercellular; (3) by centrifugal principle, manufacture a kind of weightless environment, make to all directions, to grow in cell energy three-dimensional space.
In above-mentioned cell three-dimensional training method, pass through the mode that three-dimensional rack is cultivated in conjunction with filling system, if notification number is the disclosed < < of patent document rotary pouring type bioreactor system > > of CN 1288235C, it comprises bio-reactor, this bio-reactor comprises base, base is provided with reaction vessel, in this reaction vessel, be filled with nutrient solution and be installed with three-dimensional rack, this reaction vessel injects reaction vessel by filling system by nutrient solution, the gaseous interchange inside and outside bio-reactor is carried out by semi-permeable membranes in these reaction vessel two ends.This rotary pouring type bioreactor system, can well improve cell culture environment, promote cell being more evenly distributed on three-dimensional rack, promote oxygen and nutritive substance to carry to internal stent, keep the activity of internal stent cell and the performance of function, the inside and outside gas real-time exchange of realization response device, in assurance nutrient solution, the content of oxygen and carbonic acid gas is basicly stable, maintain physiological pH value, for cellular metabolism and Function provide more favourable microenvironment.Nutrient solution in above-mentioned bio-reactor is in order to promote growth and the differentiation of cell, generally comprise common nutrient medium and the inducing culture factor, common nutrient medium generally comprises basic nutrient if sugar, amino acid, trace element etc. are to maintain growth and the propagation of cell, also comprise that damping fluid is to regulate suitable physiological PH value, the inducing culture factor plays a role as the key component of Growth of Cells differentiation, but because it is compared with difficult acquisition with yield poorly down extremely expensive.Because above-mentioned bio-reactor only has a reaction vessel, grown cultures liquid and the inducing culture factor all must be blended in same reaction vessel, the required inducing culture factor consumption of once cultivating is very large; Common nutrient medium is comparatively cheap, and for meeting Growth of Cells, nutrient solution needs regular replacing, and along with the replacing of common nutrient medium, the inducing culture factor also runs off thereupon at every turn, so cultivates, and the cost spending is higher.
Summary of the invention
In order to overcome the deficiencies in the prior art, the invention provides a kind of two-chamber dimensional perfusion bioreactor system, this two-chamber dimensional perfusion bioreactor system improves the utilization ratio of the inducing culture factor greatly by the design of double cavity structure and dialysis membrane assembly, reduce and cultivate cost, compared with dimensional perfusion formula of the prior art, cultivate more economical material benefit.
To achieve these goals, the technical solution used in the present invention is: a kind of two-chamber dimensional perfusion bioreactor system, comprise bio-reactor, in bio-reactor, be provided with three-dimensional rack, bio-reactor is connected with device for casting, it is characterized in that: described bio-reactor comprises first cavity volume of filling for common nutrient medium, and second cavity volume of filling for the inducing culture factor, between the first cavity volume and the second cavity volume, be provided with and can be less than the dialysis membrane assembly that the composition of the inducing cell factor in the inducing culture factor passes through for molecular weight, described three-dimensional rack is arranged in described the second cavity volume.
In said structure, the design of double cavity structure is placed in two chambeies common nutrient medium and the inducing culture factor, and by the dialysis membrane assembly between two-chamber, the required nutrient of growth and proliferation of cell can being exchanged at any time in two chambeies, the macromolecular substance such as the inducing cell factor in the inducing culture factor are isolated in the second cavity volume; When changing common nutrient medium, can retain the inducing culture factor, greatly reduce the consumption of the inducing cell factor, improve its effective rate of utilization, greatly reduce cultivation cost.
As the further setting of the present invention, in described three-dimensional rack, be provided with micropore or microchannel, described micropore or microchannel converge two inlet and outlet pipings of formation and are respectively artery liquid-inlet pipe and vein drain pipe, and described artery liquid-inlet pipe and vein drain pipe are connected to form respectively the internal liquid cycling mechanism of three-dimensional rack with described device for casting.
In said structure, three-dimensional rack can be provided with the micro-or microchannel of natural or artificial three-dimensional, and by this micropore or the final internal liquid cycling mechanism that forms a similar human vas circulation of microchannel, three-dimensional rack can be full of and form a full three-dimensional structure, cell can be attached on three-dimensional rack better, make the cell in each corner of internal stent obtain good gas and metabolic substd exchange, when using organ to go cell support and some to there is the man-made support of good biocompatibility, in the cell cultures later stage, can form tissue and the organ that shape is suitable, and can be transplanted in body on corresponding blood vessel by arteriovenous liquid in-out Guan Zhi, above-mentioned artery liquid-inlet pipe and vein drain pipe refer to that the liquid being similar in human vas is flow to by artery, the sanguimotor install pipeline that vein flows out.
As the further setting of the present invention, described internal liquid cycling mechanism comprises respectively and described artery liquid-inlet pipe and the needle assembly that is connected with described vein drain pipe, the other end that described needle assembly is connected with described artery liquid-inlet pipe or vein drain pipe is relatively connected with described device for casting, described needle assembly comprises at least two syringe needles, be respectively the first syringe needle and the second syringe needle, the afterbody of the first syringe needle is connected in series from beginning to end by the head of silica gel head and described the second syringe needle.
In said structure, needle assembly connects by two syringe needles, and the silica gel head of two needle support has one end that certain elasticity makes this first syringe needle be connected with artery liquid-inlet pipe and has better swing, be convenient to install three-dimensional rack and with the installation of needle assembly; Arranging of this silica gel head can be played the effect that Anti-bubble enters the second cavity volume in addition, when this silica gel head is full of after liquid, the stopping property connecting between two syringe needles is good, and after the second syringe needle and being connected of device for casting disconnect, if there is no the setting of silica gel head, airborne gas can enter in the second cavity volume thereupon, when serious, can stop up the pipe vein system system in three-dimensional rack, affect the cultivation of cell.
As the further setting of the present invention, described the second cavity volume sidewall has the through hole passing for described needle assembly, described the second cavity volume sidewall is provided with the support chamber of protruding outwardly, the head of described the first syringe needle is arranged in described the second cavity volume through described through hole, and the afterbody of described the first syringe needle is fixedly arranged on described the second cavity volume sidewall by described support chamber.
In said structure, needle assembly only has its head to be arranged in the second cavity volume, and other parts are installed in outside the second cavity volume by being fixedly arranged in the support chamber of the second cavity volume sidewall, increased in the second cavity volume can operating space, improved flexibility of operation, as operation of fixed support, connecting needle assembly and support etc.
As the further setting of the present invention, two sidewalls that described the second cavity volume is relative are respectively equipped with outer circulation import and outer circulation outlet, and described outer circulation import and outer circulation outlet are connected to form respectively the outside liquid cycling mechanism of one-way flow with described device for casting.
In said structure, this outside liquid cycling mechanism refers to and is positioned at three-dimensional rack outside, and be positioned at the liquid circulation mechanism of the second cavity volume, this circulation is a cycling mechanism that is positioned at three-dimensional rack outside of one-way flow, this outside liquid cycling mechanism can reduce the situation of cell adhesion on the second cavity volume inwall or bottom surface, cell adhered thereto is brought in three-dimensional rack by this outside liquid circulation as far as possible, allows cell cultivate in three-dimensional rack; Inner-outer circulation is used in conjunction with, and effect is better; In addition, the nutrient solution that can also carry out by controlling device for casting interval circulates, and cell in three-dimensional rack can be more easily affixed in three-dimensional rack and can not be flushed away.
As the further setting of the present invention, two sidewalls of one end that described the second cavity volume is connected with described the first cavity volume offer draw-in groove, described dialysis membrane assembly is embedded in draw-in groove, described dialysis membrane assembly comprise the first clamping plate, the second clamping plate and be attached at the first clamping plate and the second clamping plate between Mierocrystalline cellulose semi-permeable membranes.
In said structure, between the first cavity volume and the second cavity volume, be provided with rectangular communicating passage, the both sides sidewall of this communicating passage offers respectively corresponding draw-in groove in its vertical direction, this dialysis membrane assembly is embedded at the effect of playing isolation the first cavity volume and the second cavity volume in this communicating passage by draw-in groove, easy for installation, be convenient to change simultaneously; Above-mentioned the first clamping plate and the second clamping plate are the clamping plate setting with groove structure, and semi-permeable membranes is located between two clamping plate and is arranged in groove, and the good sealing effect that this structure arranges and dialysis buffer action are good.
As the further setting of the present invention, in described the second cavity volume, towards one end of described dialysis membrane assembly, be embedded with the partition that described the first cavity volume of blocking-up is communicated with the second cavity volume.
In said structure, this partition can completely cut off two cavity volumes completely, and what facilitate two cavity volumes independently changes liquid.
As the further setting of the present invention, described bio-reactor also comprises the bulb apparatus that degass, described its bubble assembly comprises liquid storage pipe, the lower end of liquid storage pipe is connected with described device for casting, the upper end of liquid storage pipe is connected with negative pressure suction device, and the sidewall of liquid storage pipe is provided with the draining hole being communicated with described the second cavity volume.
In said structure, the negative pressure suction device degassing in bulb apparatus can adopt the syringe that removes syringe needle, negative pressure suction device can will have the bubble removal of generation in the second cavity volume, guarantee that the liquid entering in the second cavity volume does not have bubble, guarantee that the pipe vein system system in three-dimensional rack can not blocked by bubble.
As the further setting of the present invention, the floorage of described the second cavity volume is 1/3 of the first cavity volume, and described the second cavity volume height is half of described the first cavity volume height.
In said structure, the vertical height of the second cavity volume is only half of the first cavity volume; Bossed window is also established in the top of above-mentioned the second cavity volume, is covered with transparent cover body on window; The present invention designs by reducing the opening of vertical height and the second cavity volume window, be very easy in operation relevant to three-dimensional rack in the second cavity volume and three-dimensional rack cell cultivation process by the cell growth condition in stereomicroscope observation three-dimensional rack, because less height degree can meet the requirement that stereoscopic microscope must be very near to viewing distance.
As the further setting of the present invention, above-mentioned bio-reactor is that transparent material is made.The transparent design of two cavity volumes, has facilitated the light of bottom to be shining into requirement, has also facilitated fluorescence real time imagery to observe simultaneously.
Adopt such scheme, the present invention can allow cell in system, obtain good nutrient and gaseous interchange by two-chamber dimensional perfusion bioreactor system, refuse can well be discharged, and under the effect of common nutrient medium and the inducing culture factor, better growing multiplication and differentiation become object cell to cell; And the design of the double cavity structure of bio-reactor can be so that the common nutrient medium in two cavity volumes and the inducing culture factor be independently changed liquid, when changing common nutrient medium, can retain the inducing culture factor, greatly reduce the consumption of the inducing cell factor, improve its effective rate of utilization, reduce costs.
Below in conjunction with accompanying drawing, the invention will be further described.
Accompanying drawing explanation
Accompanying drawing 1 is specific embodiment of the invention structural representation;
Accompanying drawing 2 is specific embodiment of the invention structural front view;
Accompanying drawing 3 is the A-A sectional view of specific embodiment of the invention accompanying drawing 2;
The structure sectional view of the bio-reactor that accompanying drawing 4 is the specific embodiment of the invention;
Accompanying drawing 5 is specific embodiment of the invention structure right view;
Accompanying drawing 6 is specific embodiment of the invention structure left view;
The structural representation of accompanying drawing 8 specific embodiment of the invention needle assemblies;
Accompanying drawing 9 is the internal structure schematic diagram of the communicating passage in specific embodiment of the invention accompanying drawing 4;
Accompanying drawing 10 is the another kind of internal structure schematic diagram of the communicating passage in specific embodiment of the invention accompanying drawing 4.
Embodiment
Specific embodiments of the invention are two-chamber dimensional perfusion bioreactor systems as shown in Fig. 1-9, comprise bio-reactor 1, in bio-reactor 1, be provided with three-dimensional rack 2, bio-reactor 1 is connected with device for casting, bio-reactor comprises base 3, first cavity volume 11 of filling for common nutrient medium, and second cavity volume 12 of filling for the inducing culture factor, the first cavity volume 11 and the second cavity volume 12 are positioned on base 3, and base 3 plays the effect of peristaltic pump of the power resources of support and fixed cycles device for casting; Between the first cavity volume 11 and the second cavity volume 12, be provided with and can be less than the dialysis membrane assembly 4 that the composition of the inducing cell factor in the inducing culture factor passes through for molecular weight, three-dimensional rack 2 is arranged in the second cavity volume 12.The design of double cavity structure is placed in two chambeies common nutrient medium and the inducing culture factor, and by the dialysis membrane assembly 4 between two-chamber, the required nutrient of growth and proliferation of cell can being exchanged at any time in two chambeies, the macromolecular substance such as the inducing cell factor in the inducing culture factor are isolated in the second cavity volume 12; When changing common nutrient medium, can retain the inducing culture factor, greatly reduce the consumption of the inducing cell factor, improve its effective rate of utilization, greatly reduce cultivation cost.
In above-mentioned three-dimensional rack 2, be provided with artificial or natural three-dimensional micropore or microchannel, micropore or microchannel converge two inlet and outlet pipings of formation and are respectively artery liquid-inlet pipe and vein drain pipe, and artery liquid-inlet pipe and vein drain pipe are connected to form respectively the internal liquid cycling mechanism of three-dimensional rack 4 with device for casting.The range of choice of above-mentioned three-dimensional rack 2 is very wide, what can be various histoorgans as liver, lungs, heart, kidney removes cell support, removing cytoskeleton is by removing organ and in-house original cell, retaining the three-dimensional rack 2 that extracellular matrix skeleton forms.This three-dimensional rack 2 has the structure that inner vascular canal etc. meets normal physiological, and by being connected into above-mentioned internal liquid cycling mechanism part, the Growth of Cells three-dimensional environment in just can analogue body, reaches the growth and the Differentiation that promote stem cell.Also can select some thicker synthetic supports, as long as possess the circulation path of above-mentioned similar sanguimotor turnover, all can apply, same organ 3D print carriage also can be applied.
In said structure, in three-dimensional rack 2, form the internal liquid cycling mechanism of a similar human vas circulation, three-dimensional rack 2 can be full of and form a full three-dimensional structure, make cell can be attached to better on three-dimensional rack 2, make the cell in three-dimensional rack 2 inner each corners obtain good gas and metabolic substd exchange, when using organ to go cell support and some to there is the man-made support of good biocompatibility, in the cell cultures later stage, can form tissue and the organ that shape is suitable, and can be transplanted in body on corresponding blood vessel by arteriovenous liquid in-out Guan Zhi, above-mentioned artery liquid-inlet pipe and vein drain pipe refer to that the liquid being similar in human vas is flow to by artery, the sanguimotor install pipeline that vein flows out.
Above-mentioned device for casting comprises peristaltic pump 51 and pump line 52, the mouth of pipe deck that the base 3 of above-mentioned bio-reactor 1 is provided with corresponding peristaltic pump deck and coordinates with pump line 52, is installed on peristaltic pump 51 on base 3, forms an integral body, coordinate compactness, reduce and take up an area space.
As shown in Fig. 3,8, above-mentioned internal liquid cycling mechanism comprises respectively and artery liquid-inlet pipe and the needle assembly 6 that is connected with vein drain pipe, the other end that needle assembly 6 is connected with artery liquid-inlet pipe or vein drain pipe is relatively connected with pump line 52, needle assembly 6 comprises two syringe needles, the afterbody that is respectively the first syringe needle 61 and the second syringe needle 62, the first syringe needles 61 is connected in series from beginning to end by the head of silica gel head 63 and the second syringe needle 62.Needle assembly 6 connects by two syringe needles, and the silica gel head 63 between two syringe needles has one end that certain elasticity makes this first syringe needle 61 be connected with artery liquid-inlet pipe and has better swing, be convenient to install three-dimensional rack 2 and with the installation of needle assembly 6; Arranging of this silica gel head 63 can be played the effect that Anti-bubble enters the second cavity volume 12 in addition, when this silica gel head 63 is full of after liquid, the stopping property connecting between two syringe needles is good, and after the second syringe needle 62 and being connected of device for casting disconnect, if there is no the setting of silica gel head 63, airborne gas can enter in the second cavity volume 12 thereupon, when serious, can stop up the pipe vein system system in three-dimensional rack 2, affect the cultivation of cell.
Above-mentioned the second cavity volume 12 sidewalls have for two two through holes that needle assembly 6 passes, the second cavity volume 12 sidewalls are provided with the support chamber 121 of protruding outwardly, this support chamber 121 by bracket around forming, the afterbody that the head of the first syringe needle 61 is arranged in the second cavity volume 12, the first syringe needles 61 through through hole is fixedly arranged on the second cavity volume 12 sidewalls by support chamber 121.Needle assembly 6 only has its head to be arranged in the second cavity volume 12, and other parts are installed in outside the second cavity volume 12 by being fixedly arranged on the support chamber 121 of the second cavity volume 12 sidewalls, increased in the second cavity volume 12 can operating space, improved flexibility of operation, as the operation of fixing three-dimensional rack 2, connecting needle assembly 6 and three-dimensional rack 2 etc.
Two sidewalls that above-mentioned the second cavity volume 12 is relative are respectively equipped with the interface 13 being connected with outer circulation outlet with outer circulation import, and the interface 13 of the interface 13 of outer circulation import and outer circulation outlet is connected to form respectively the outside liquid cycling mechanism of one-way flow with device for casting.This outside liquid cycling mechanism refers to and is positioned at three-dimensional rack 2 outsides, and be positioned at the liquid circulation mechanism of the second cavity volume 12, this circulation is a cycling mechanism of perforation three-dimensional rack 2 outsides of one-way flow, this outside liquid cycling mechanism can reduce the situation of cell adhesion on the second cavity volume 12 inwalls or bottom surface, cell adhered thereto is brought in three-dimensional rack 2 by this outside liquid circulation as far as possible, allows cell cultivate in three-dimensional rack 2; Inner-outer circulation is used in conjunction with, and effect is better; In addition, can also carry out the circulation at interval by controlling device for casting, cell in three-dimensional rack 2 can be more easily affixed in three-dimensional rack 2 and can not be flushed away.
As Fig. 3, 4, shown in 9, between above-mentioned the second cavity volume 12 and the first cavity volume 11, be provided with rectangular communicating passage 14, the both sides sidewall of this communicating passage 14 offers respectively corresponding draw-in groove 141 in its vertical direction, dialysis membrane assembly 4 is embedded in draw-in groove 141, dialysis membrane assembly 4 comprises the first clamping plate 41, the second clamping plate 42, and be attached at the Mierocrystalline cellulose semi-permeable membranes 43 between the first clamping plate 41 and the second clamping plate 42, the general inducing cell factor is all macro-molecular protein, therefore this semi-permeable membranes 43 is generally less than 3.5kd, in specific experiment, can arrange according to the size of the different inducing cell factor protein molecular weight in the inducing culture factor.This dialysis membrane assembly 4 is embedded in communicating passage 14 by draw-in groove 141, easy for installation, is convenient to change simultaneously; Above-mentioned the first clamping plate 41 and the second clamping plate 42 are the clamping plate setting with groove 44 structures, and semi-permeable membranes 43 is located between two clamping plate and is arranged in groove 44, and the good sealing effect that this structure arranges and dialysis buffer action are good.
As shown in Figure 4, in above-mentioned the second cavity volume 12, towards one end of dialysis membrane assembly 4, be embedded with the partition 7 that blocking-up the first cavity volume 11 is communicated with the second cavity volume 12.This partition 7 can completely cut off two cavity volumes completely, and what facilitate two cavity volumes independently changes liquid.Above-mentioned communicating passage 14 sidewalls are also provided with the groove 142 inserting for this partition 7, this groove 142 leaves space at above-mentioned draw-in groove 141 between one end of the second cavity 12 and partition 7 and dialysis membrane assembly 4, and both are for being independently installed in communicating passage 14 as shown in figure 10.Independent dialysis membrane assembly 4 and the dividing plate 7 of installing has better sealing property.
As shown in Figure 1, above-mentioned bio-reactor 1 also comprises the bulb apparatus 8 that degass, its bubble assembly 8 comprises liquid storage pipe 81, the lower end of liquid storage pipe 81 with by Medical tee joint pipe, be connected with pump line 52, the upper end of liquid storage pipe 81 is connected with negative pressure suction device 82, and the sidewall of liquid storage pipe 81 is provided with the draining hole 811 that the needle assembly 6 that is connected with artery liquid-inlet pipe on three-dimensional rack 2 is communicated with.The negative pressure suction device 82 degassing in bulb apparatus 8 can adopt the syringe that removes syringe needle, negative pressure suction device 82 can will have the bubble removal of generation in the second cavity volume 12, guarantee that the liquid entering in the second cavity volume 12 does not have bubble, guarantee that the pipe vein system system in three-dimensional rack 2 can not blocked by bubble.
The floorage of above-mentioned the second cavity volume 12 is that 1/3, the second cavity volume 12 of the first cavity volume 11 is highly half of the first cavity volume 11 height.The vertical height of the second cavity volume 12 is only half of the first cavity volume 11; Bossed window 122 is also established in the top of above-mentioned the second cavity volume 12, is covered with transparent cover body 123 on window 122; The present invention is by reducing the vertical height of the second cavity volume 12, and window 13 designs of the second cavity volume 12, be very easy in operation relevant to three-dimensional rack 2 in the second cavity volume 12 and three-dimensional rack 2 cell cultivation process by the cell growth condition in stereomicroscope observation three-dimensional rack, because less height degree can meet the requirement that stereoscopic microscope must be very near to viewing distance.
As shown in Figure 3, the other end that above-mentioned the first cavity volume 11 is connected with the second cavity volume 12 is relatively provided with pipe connecting 111, on this pipe connecting 111, be arranged with sealing-ring 114, the access flap 112 that has been threaded on pipe connecting 111, offers the standard interface 113 being connected with device for casting on access flap 112.In the present invention, relate to the interface or the pipeline that are connected with device for casting and be standardized component setting, be convenient to configuration.The first cavity volume 11 is by pipe connecting 111 designs of openable, liquid light in cavity changed places and inject or remove, and in addition, also reduced the possibility of the bacterial contamination in culturing process.This design not only can feed liquor body also can air inlet body, by bubbling style gas, in conjunction with sending out, the bubble that can well avoid culture apparatus cell to be produced by bubbling device damages.
Above-mentioned bio-reactor 1 is made for transparent material, according to different working conditions, can be made by optically transparent polystyrene and other polyethylene materials.The transparent design of two cavity volumes, has facilitated the light of bottom to be shining into requirement, has also facilitated fluorescence real time imagery to observe simultaneously.
Adopt such scheme, the present invention can allow cell in system, obtain good nutrient and gaseous interchange by two-chamber dimensional perfusion bioreactor system, refuse can well be discharged, and under the effect of common nutrient medium and the inducing culture factor, better growing multiplication and differentiation become object cell to cell; And the design of the double cavity structure of bio-reactor 1 can be so that the common nutrient medium in two cavity volumes and the inducing culture factor be independently changed liquid, when changing common nutrient medium, can retain the inducing culture factor, greatly reduce the consumption of the inducing cell factor, improve its effective rate of utilization, reduce costs.
Claims (10)
1. a two-chamber dimensional perfusion bioreactor system, comprise bio-reactor, in bio-reactor, be provided with three-dimensional rack, bio-reactor is connected with device for casting, it is characterized in that: described bio-reactor comprises first cavity volume of filling for common nutrient medium, and second cavity volume of filling for the inducing culture factor, between the first cavity volume and the second cavity volume, be provided with and can be less than the dialysis membrane assembly that the composition of the inducing cell factor in the inducing culture factor passes through for molecular weight, described three-dimensional rack is arranged in described the second cavity volume.
2. two-chamber dimensional perfusion bioreactor system according to claim 1, it is characterized in that: in described three-dimensional rack, be provided with micropore or microchannel, described micropore or microchannel converge two inlet and outlet pipings of formation and are respectively artery liquid-inlet pipe and vein drain pipe, and described artery liquid-inlet pipe and vein drain pipe are connected to form respectively the internal liquid cycling mechanism of three-dimensional rack with described device for casting.
3. two-chamber dimensional perfusion bioreactor system according to claim 2, it is characterized in that: described internal liquid cycling mechanism comprises respectively and described artery liquid-inlet pipe and the needle assembly that is connected with described vein drain pipe, the other end that described needle assembly is connected with described artery liquid-inlet pipe or vein drain pipe is relatively connected with described device for casting, described needle assembly comprises at least two syringe needles, be respectively the first syringe needle and the second syringe needle, the afterbody of the first syringe needle is connected in series from beginning to end by the head of silica gel head and described the second syringe needle.
4. two-chamber dimensional perfusion bioreactor system according to claim 3, it is characterized in that: described the second cavity volume sidewall has the through hole passing for described needle assembly, described the second cavity volume sidewall is provided with the support chamber of protruding outwardly, the head of described the first syringe needle is arranged in described the second cavity volume through described through hole, and the afterbody of described the first syringe needle is fixedly arranged on described the second cavity volume sidewall by described support chamber.
5. according to the two-chamber dimensional perfusion bioreactor system described in claim 1-4 any one, it is characterized in that: two sidewalls that described the second cavity volume is relative are respectively equipped with outer circulation import and outer circulation outlet, described outer circulation import and outer circulation outlet are connected to form respectively the outside liquid cycling mechanism of one-way flow with described device for casting.
6. two-chamber dimensional perfusion bioreactor system according to claim 1, it is characterized in that: two sidewalls of one end that described the second cavity volume is connected with described the first cavity volume offer draw-in groove, described dialysis membrane assembly is embedded in draw-in groove, described dialysis membrane assembly comprise the first clamping plate, the second clamping plate and be attached at the first clamping plate and the second clamping plate between Mierocrystalline cellulose semi-permeable membranes.
7. according to the two-chamber dimensional perfusion bioreactor system described in claim 1 or 6, it is characterized in that: in described the second cavity volume, towards one end of described dialysis membrane assembly, be embedded with the partition that described the first cavity volume of blocking-up is communicated with the second cavity volume.
8. two-chamber dimensional perfusion bioreactor system according to claim 1, it is characterized in that: described bio-reactor also comprises the bulb apparatus that degass, described its bubble assembly comprises liquid storage pipe, the lower end of liquid storage pipe is connected with described device for casting, the upper end of liquid storage pipe is connected with negative pressure suction device, and the sidewall of liquid storage pipe is provided with the draining hole being communicated with described the second cavity volume.
9. two-chamber dimensional perfusion bioreactor system according to claim 1, is characterized in that: the floorage of described the second cavity volume is 1/3 of the first cavity volume, and described the second cavity volume height is half of described the first cavity volume height.
10. two-chamber dimensional perfusion bioreactor system according to claim 1, is characterized in that: described bio-reactor is that transparent material is made.
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