CN105012050A - Method and special mould for preparing tissue and organ precursor with multi-branch channels - Google Patents
Method and special mould for preparing tissue and organ precursor with multi-branch channels Download PDFInfo
- Publication number
- CN105012050A CN105012050A CN201510419730.XA CN201510419730A CN105012050A CN 105012050 A CN105012050 A CN 105012050A CN 201510419730 A CN201510419730 A CN 201510419730A CN 105012050 A CN105012050 A CN 105012050A
- Authority
- CN
- China
- Prior art keywords
- outer mold
- mould
- level
- mold
- layer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Prostheses (AREA)
Abstract
The invention relates to a method and special mould for preparing a tissue and organ precursor with multi-branch channels, wherein the special mould comprises a base mould, at least one stage of inner mould and at least one stage of outer mould. The method provided by the invention comprises the steps that: the inner mould and the outer mould are arranged on the base mould in sequence according to the stages, when each stages of mould is mounted, a matrix solution containing cells is poured in the outer mould cavity, after the crosslinking of the matrix solution containing cells, the outer mould and the inner mould are removed, so that a layer of cell matrix layer with channels is formed; then the above steps are repeated, so that multiple layers of the cell matrix layer with channels are prepared in sequence, the interlayer channels are interconnected in branch-shaped, the multilayer cell matrix layer is penetrated and adhered through the synthetic polymer solution to form the tissue and organ precursor with multi-branch channels, and the shaped multi-branch channels can form the structures, such as blood vessel, nervus, bile duct, trachea and the like. The method provided by the invention overcome a defect that it is difficult to construct the complex branch channel in prior technology, and has advantages including that: the range of shaped tissue and organ is wide, the mounting of matrix materials containing high density of cells is easy to be realize.
Description
Technical field
The invention belongs to the artificial manufacturing technology field of organism complicated tissue organ, particularly a kind of method and particular manufacturing craft preparing the histoorgan precursor of band multiple-limb passage.
Background technology
The patient suffering from tissue defect or organ failure in the world every year exceedes ten million, so, the reparation of human body disease damage tissue and organ and alternative be a sciences problems needing solution badly, be also the science frontier attracted people's attention this century.The intrinsic drawback such as to have a big risk because biological tissue organ transplantation exists donor shortage and immunologic rejection, be difficult to become the effective way solving this problem.Accordingly, arise at the historic moment by the organizational project (Tissue Engineering) improved for the purpose of this type of illness treatment level, organizational project is the principle of integrated application biology, materialogy and engineering, build the three-dimensional structure of cell and material in vitro, by developing into the transplantation substitute had with natural tissues similar structures and function after suitable cultivation and training.The scientific meaning important due to it and potential economic worth, organizational project receives the great attention of countries in the world always, obtains the development of advancing by leaps and bounds, and achieve many achievements short less than in the time of 30 years.But with regard to the application of its product, mainly concentrate on relatively simple a few field such as bone tissue engineer and skin tissue engineering of structure at present.Trace it to its cause, the very important point is exactly, the vitro construction method of existing organizational project structure is (as Electrospun, 3D impact system, molten modulus method etc.) be difficult to build the blood vessel network structure with the similar complexity of natural tissues, and blood vessel is the part that must not lack in histoorgan growth, can be histoorgan growth and necessary oxygen and nutrient substance are provided, research shows, cell only around blood vessel within the scope of 150-200 μm can obtain nutrient and oxygen (Colton C.Implantable biohybrid artificialorgans.Cell Translant by dispersion, 1955, 4:415-436), lack complicated blood vessel network and cause the cell survival rate of construction inner low, be therefore difficult to the structure realizing bulk tissue.In organizational project, the method of conventional structure blood vessel mainly contains and relies on somatomedin slow releasing to promote vascularization, many cells Combined culture forms blood vessel structure, utilizes bioreactor and microfluidic channel to form blood vessel and the vascularization etc. based on advanced forming technique.
In addition, the structure that human body is made up of Various Tissues and organ, the function of the physiological process in body and various histoorgan is all generally directly and under indirect adjustments and controls mutually carry out neural, isolated organ can not realize the function on biological significance, and nervous system is made in rapid and perfect adjustment in the change of human body to inside and outside environment and played conclusive effect.So when structure has the human organ of specific function in vitro, inevitably introduce nervous system.Similar with vascularity, nerve is also the net distribution according to multilevel branch in human body.In addition, bile duct, trachea and urinary catheter etc. also play an important role in human body, so the histoorgan precursor built containing multiple-limb passage is key one step realizing histoorgan biological function in vitro.
Patent of invention document (application number 201110448154.3) provide a kind of utilize the layering of assembling die pour into be shaped containing the method for the complicated tissue organ precursor of multiple-limb blood vessel structure.Infusion to containing in the intermediate mold of different-diameter between outer mold, is removed mould and is namely obtained histoorgan precursor by this invention after crosslinking Treatment.The shortcoming of this invention is that the structure of internal blood vessel depends on the structure of intermediate mold completely, therefore can only the simple blood vessel structure of molding structure, for building, branch vessel web frame complicated in natural tissues is helpless, in addition, the method utilizes annular intermediate mold to wrap up the forming method of inner mold, assembly manipulation difficulty is large, inevitably damages in type structure in unloading process.
Patent of invention (application number 201210324600.4) proposes a kind of method utilizing rotation combination mould to prepare fusiform complex organ precursor.This invention utilizes the Weissenbery effect of non-Newtonian fluid, the fusiform complex organ precursor of band multiple-limb blood vessel can be prepared in theory, but the shortcoming of the method is the rotary speed of mould and the climb altitude of liquid does not have clear and definite quantitative relationship, the height determining internal ring mould is difficult to during practical operation, and shaped structure is single, can only formed profile be fusoid histoorgan.In addition, each shaping all defines the spindle of band cone point, therefore during forming belt multilevel branch blood vessel structure, the second level and above internal ring mould thereof are difficult to location and installation on next stage fulcrum, and the organ shape that the method is shaped in addition can only be single fusiform.
Summary of the invention
The object of this invention is to provide a kind of method and the particular manufacturing craft of preparing the histoorgan precursor of band multiple-limb passage, be intended on the basis of work on hand, utilize the layering of particular manufacturing craft assembling way to be shaped and there is the histoorgan of branched bottom, to overcome the drawback that prior art is difficult to have in histoorgan internal build complex branches passage, avoid complicated assembly manipulation simultaneously.
Technical scheme of the present invention is as follows:
Prepare a particular manufacturing craft for the histoorgan precursor of band multiple-limb passage, it is characterized in that: described particular manufacturing craft comprises base mould, at least one-level outer mold and at least one-level inner mold; Described base mould inside is for becoming large stepped from bottom to up successively, and step number is identical with outer mold progression, and base mould bottom surface is plane, concave surface or convex surface, and is distributed with locating hole; Every grade of outer mold is equipped with installing hole and perfusing hole, the ladder of the external dimensions of outer mold and the corresponding progression of base mould to surround the size in face identical, the inner chamber of next stage outer mold and the inner chamber of upper level outer mold seamlessly transit, and the inner chamber of outer mold has the profile identical with presetting histoorgan, the centrage of the perfusing hole of multistage outer mold is positioned at same straight line; Every grade of inner mold comprises at least one branch-like structure, its number is identical with the number of the installing hole of respective stages outer mold, each branch-like structure comprises total arm and branched pipe, total arm of each branch-like structure and the installing hole of respective stages outer mold form matching relationship, the number of the branched pipe of most higher level's inner mold is 1, and total manifold size of next stage inner mold is identical with the branched pipe size of adjacent upper level inner mold.
Described a kind of particular manufacturing craft preparing the histoorgan precursor of band multiple-limb passage, is characterized in that: the material of described base mould and outer mold is hard synthesized polymer material or metal material; Inner mold is the solid tubulose be made up of inanimate object toxic materials, and concrete material is including, but not limited to synthetic fibers, polyethylene, Merlon, polrvinyl chloride and polyurethane.
Utilize particular manufacturing craft to prepare the method for the histoorgan precursor of band multiple-limb passage, it is characterized in that the method comprises the steps:
1) preparation quality volumetric concentration be the hydrogel solution of 1% ~ 30% as matrix solution, extract or buy zooblast make cell suspending liquid, cell suspending liquid density is 1 × 10
6~ 5 × 10
7individual/ml, is mixed and made into celliferous matrix solution in 1 ~ 9:9 ~ 1 by volume by matrix solution and cell suspending liquid, is dissolved in by synthesized polymer material in organic solvent and makes the synthesis macromolecular solution that mass percentage concentration is 1% ~ 30%;
2) three-dimensional printing technology is utilized to prepare particular manufacturing craft, total arm of first order inner mold is arranged on the installing hole of first order outer mold, first order outer mold is enclosed within the first order step of base mould, first order inner mold branched pipe is arranged in the locating hole of base mould bottom surface, celliferous matrix solution is poured into mould from the perfusing hole of first order outer mold, physics or chemical crosslinking or polymerization is adopted after filling first order outer mold and base mould gap, celliferous matrix solution in mould is cross-linked, first order outer mold and first order inner mold is removed successively after being cross-linked, form stable ground floor cellular matrix layer, wherein containing the first order branched bottom formed by first order inner mold,
3) total arm of second level inner mold is arranged on the installing hole of second level outer mold, second level outer mold is enclosed within the second level step of base mould, the branched pipe of second level inner mold aims at the passage on the ground floor cellular matrix layer that has been shaped, celliferous matrix solution is poured into mould from the perfusing hole of second level outer mold, physics or chemical crosslinking or polymerization is adopted after filling second level outer mold and ground floor cellular matrix lamellar spacing, celliferous matrix solution is cross-linked, second level outer mold and second level inner mold is removed successively after being cross-linked, form the stable second layer cellular matrix layer with second level passage,
4) step 3 is repeated), form third layer respectively to the cellular matrix layer of the superiors, obtain the multi-layer cellular hypothallus of the mutually through band multiple-limb passage of inter-layer passages;
5) synthesis macromolecular solution is sprayed onto multi-layer cellular substrate layer surface formed thereby, after extraction, forms synthesis macromolecule layer, remove base mould, namely form the histoorgan precursor of band multiple-limb passage.
The preparation method of the histoorgan precursor of described band multiple-limb passage, is characterized in that: the passage in the histoorgan precursor of band multiple-limb passage forms blood vessel, nerve, bile duct, trachea or urinary catheter in histoorgan precursor incubation.
The preparation method of the histoorgan precursor of described band multiple-limb passage, it is characterized in that: the concrete grammar described passage being formed as blood vessel is, preparation is containing the matrix solution of endotheliocyte, first the matrix solution of one deck containing endotheliocyte is applied at mold surface before being shaped, adopt physics or chemical crosslinking or polymerization, matrix solution containing endotheliocyte is cross-linked, spray endotheliocyte suspension and synthesis macromolecular solution subsequently successively, hypothallus-endothelial layer-synthesis macromolecule layer the three-decker containing endotheliocyte is formed at mold surface after extraction, shaping completes cellular matrix layer and carefully extracts inner mold, this three-decker is stayed passage surrounding.
The preparation method of the histoorgan precursor of described band multiple-limb passage, is characterized in that: described passage being formed as neural concrete grammar is, the matrix solution containing Schwann cell, containing the matrix solution of Schwann cell, is directly poured into stand in channel by preparation.
The preparation method of the histoorgan precursor of described band multiple-limb passage, is characterized in that: described hydrogel is one or more in gelatin, Fibrinogen, collagen, chitosan, sodium alginate, hyaluronic acid and fibronectin.
The preparation method of the histoorgan precursor of described band multiple-limb passage, it is characterized in that: described synthesized polymer material adopts the complex of one or more materials in polyurethane, polycaprolactone, Merlon, polylactic acid, Polyethylene Glycol, polylactic acid, polyester, polyhydroxy acid ester and lactic acid and ethanol copolymer, organic solvent for dissolving synthesized polymer material adopts TEG, ethylene glycol, isopropyl alcohol or Isosorbide-5-Nitrae-dioxane.
The preparation method of the histoorgan precursor of described band multiple-limb passage, it is characterized in that: in described celliferous matrix solution and synthesis macromolecular solution, add one or more in endothelial cell growth factor (ECGF), cell transfer factor or hepatocyte growth factor, and one or more in heparin, paclitaxel or sulfated chitosan.
The present invention is compared to prior art, the present invention adopts assembling die layering to manufacture histoorgan precursor, the histoorgan of complexity is divided into multilamellar, often walk the cellular matrix layer of shaping one deck with passage, multistep superposition forms the histoorgan precursor with multi-layer cellular hypothallus, passage in cellular matrix layer is mutually through, in the branch-like of class natural structure; The present invention is easy and simple to handle, and die assembly is simple with dismounting.In addition, present invention uses three-dimensional printing technology to prepare particular manufacturing craft, thus the simple structure that dies cavity is not limited to simple revolving body or is made up of plane, can manufacture by the complex-curved structure formed arbitrarily in theory, expand the scope that histoorgan manufactures, the histoorgan making preparation have personalized profile becomes possibility.
Accompanying drawing explanation
Fig. 1 is the overall profile of particular manufacturing craft.
Fig. 2 a is the top view (for three stage rank) of base mould.
Fig. 2 b is the A-A sectional view of Fig. 2 a.
Fig. 3 is inner mold schematic diagram (branching into example with three).
Fig. 4 is the schematic diagram of histoorgan forming precursor process.
Fig. 5 is the histoorgan precursor profile (to be with three grades of branched bottoms) of band multiple-limb passage.
Fig. 6 is containing endotheliocyte hypothallus-endothelial layer-synthesis macromolecule layer three-decker schematic diagram.
In figure: 101-outer mold; 102-base mould; 103-inner mold; The total arm of 301-; 302-branched pipe; 401-cellular matrix layer; 402-perfusing hole; The celliferous substrate of 403-; 404-passage; 501-synthesizes macromolecule layer; 601-is containing endotheliocyte hypothallus; 602-endothelial layer.
Detailed description of the invention
In order to understand technical scheme of the present invention further, to develop simultaneously embodiment referring to accompanying drawing, the present invention is described in further detail.
Fig. 1 is the overall profile of particular manufacturing craft provided by the invention; Fig. 2 a is the top view (for three stage rank) of base mould; Fig. 2 b is the A-A sectional view of Fig. 2 a; Fig. 3 is inner mold schematic diagram; The particular manufacturing craft of the histoorgan precursor of preparation band multiple-limb passage comprises base mould 102, at least one-level outer mold 101 and at least one-level inner mold 103, outer mold and inner mold are followed successively by first order inner mold from bottom to up, first order outer mold, second level inner mold, second level outer mold,, most higher level's inner mold, most higher level's outer mold; Described base mould inside is for becoming large stepped from bottom to up successively, and step number is identical with outer mold progression, and base mould bottom surface is plane, concave surface or convex surface, and is distributed with locating hole; Every grade of outer mold is equipped with installing hole and perfusing hole 402, the ladder of the external dimensions of outer mold and the corresponding progression of base mould to surround the size in face identical, the inner chamber of next stage outer mold and the inner chamber of upper level outer mold seamlessly transit, and the inner chamber of outer mold has the profile identical with presetting histoorgan, the centrage of the perfusing hole 402 of multistage outer mold is positioned at same straight line; Every grade of inner mold comprises at least one branch-like structure, its number is identical with the number of the installing hole of respective stages outer mold, each branch-like structure comprises total arm 301 and branched pipe 302, total arm of each branch-like structure and the installing hole of respective stages outer mold form matching relationship, the number of the branched pipe of most higher level's inner mold is 1, and total manifold size of next stage inner mold is identical with the branched pipe size of adjacent upper level inner mold; The material of base mould and outer mold is hard synthesized polymer material or metal material; Inner mold is the solid tubulose be made up of inanimate object toxic materials, and concrete material is including, but not limited to synthetic fibers, polyethylene, Merlon, polrvinyl chloride and polyurethane.
Fig. 4 is the schematic diagram of histoorgan forming precursor process, and the histoorgan precursor of preparation band multiple-limb passage specifically comprises the steps:
1) preparation quality volumetric concentration be the hydrogel solution of 1% ~ 30% as matrix solution, extract or buy zooblast make cell suspending liquid, cell suspending liquid density is 1 × 10
6~ 5 × 10
7individual/ml, is mixed and made into celliferous matrix solution in 1 ~ 9:9 ~ 1 by volume by matrix solution and cell suspending liquid, is dissolved in by synthesized polymer material in organic solvent and makes the synthesis macromolecular solution that mass percentage concentration is 1% ~ 30%; The anticoagulin of one or more in one or more and heparin, paclitaxel or the sulfated chitosan in endothelial cell growth factor (ECGF), cell transfer factor or hepatocyte growth factor is optionally added in described celliferous matrix solution and synthesis macromolecular solution.
2) three-dimensional printing technology is utilized to prepare particular manufacturing craft, total arm of first order inner mold is arranged on the installing hole of first order outer mold, first order outer mold is enclosed within the first order step of base mould, first order inner mold branched pipe is arranged in the locating hole of base mould bottom surface, celliferous matrix solution is poured into mould from the perfusing hole of first order outer mold, physics or chemical crosslinking or polymerization is adopted after filling first order outer mold and base mould gap, celliferous matrix solution in mould is cross-linked, first order outer mold and first order inner mold is removed successively after being cross-linked, form stable ground floor cellular matrix layer, wherein containing the first order branched bottom formed by first order inner mold,
3) total arm of second level inner mold is arranged on the installing hole of second level outer mold, second level outer mold is enclosed within the second level step of base mould, the branched pipe of second level inner mold aims at the passage on the ground floor cellular matrix layer that has been shaped, celliferous matrix solution is poured into mould from the perfusing hole of second level outer mold, physics or chemical crosslinking or polymerization is adopted after filling second level outer mold and ground floor cellular matrix lamellar spacing, celliferous matrix solution is cross-linked, second level outer mold and second level inner mold is removed successively after being cross-linked, form the stable second layer cellular matrix layer with second level passage,
4) step 3 is repeated), form third layer respectively to the cellular matrix layer of the superiors, obtain the multi-layer cellular hypothallus of the mutually through band multiple-limb passage of inter-layer passages;
5) synthesis macromolecular solution is sprayed onto multi-layer cellular substrate layer surface formed thereby, synthesis macromolecule layer 501 is formed after extraction, remove base mould, namely form the histoorgan precursor of band multiple-limb passage, as shown in Figure 5 (to be with three grades of branched bottoms).
Preferred version of the present invention selects one or more hydrogels in gelatin, Fibrinogen, collagen, chitosan, sodium alginate, hyaluronic acid and fibronectin to be dissolved in normal saline, PBS solution, the 0.09M sodium chloride of pH=6 ~ 8,3-hydroxymethyl aminomethane hydrochloric acid solution or cell culture fluid as matrix solution; Synthesized polymer material adopts the complex of one or more materials in polyurethane, polycaprolactone, Merlon, polylactic acid, Polyethylene Glycol, polylactic acid, polyester, polyhydroxy acid ester and lactic acid and ethanol copolymer, organic solvent for dissolving synthesized polymer material adopts TEG, ethylene glycol, isopropyl alcohol or Isosorbide-5-Nitrae-dioxane; One or more cell growth factor in endothelial cell growth factor (ECGF), cell transfer factor or hepatocyte growth factor and one or more anticoagulins in heparin, paclitaxel or sulfated chitosan are added in celliferous matrix solution and synthesis macromolecular solution;
The histoorgan precursor of the band multiple-limb passage made by the present invention, described passage can be induced as blood vessel, nerve, or directly as bile duct, trachea, urinary catheter, the concrete grammar being blood vessel by described passage induction is, first at the matrix solution of mold surface coating containing endotheliocyte before being shaped, adopt physics or chemical crosslinking or polymerization, be cross-linked to form containing endotheliocyte hypothallus 601 to the matrix solution containing endotheliocyte, spray endotheliocyte suspension subsequently successively and form endothelial layer 602 and synthesis macromolecular solution, formed containing endotheliocyte hypothallus-endothelial layer-synthesis macromolecule layer three-decker (shown in Fig. 6) at mold surface after extraction, shaping completes cellular matrix layer and carefully extracts inner mold, this three-decker is stayed passage surrounding, being neural concrete grammar by described passage induction is, directly by the matrix solution perfusion stand in channel containing Schwann cell,
Embodiment 1: preparation band Pyatyi multiple-limb blood vessel and neural liver precursor
1) base mould and the Pyatyi outer mold of Pyatyi step is contained with three-dimensional printing technology preparation, dies cavity is class liver curved surface, material therefor is silicone rubber, polythene material is used to prepare Pyatyi inner mold respectively, first order inner mold contains 1 branched pipe, and all the other inner molds all contain 3 branched pipes;
2) prepare 5% fibrinogen solution as matrix solution, 10IU/mL thrombin solution, buy endotheliocyte, Schwann cell and hepatocyte and make cell suspending liquid, cell density is 1 × 10
7individual/ml, matrix solution and cell suspending liquid are mixed and made into celliferous matrix solution by 4:6 volume ratio respectively, be equipped with concentration be PLGA/ TEG (Tetraglycol) solution of 10% (w/v) as synthesis macromolecular solution, and add the heparin of 1% (w/v);
3) vascularization process is carried out to part inner mold and make Ink vessel transfusing mould, concrete grammar applies the matrix solution of one deck containing endotheliocyte at mold surface, and be cross-linked with thrombin solution, spray endotheliocyte suspension subsequently and form endothelial layer, then synthesize macromolecular solution in endothelial layer surface sprinkling and extract, so far form the hypothallus-endothelial layer-synthesis macromolecule layer three-decker containing endotheliocyte at mold surface, all the other inner molds do not process as neural inner mold;
4) total arm of first order inner mold is arranged on the installing hole of first order outer mold, in the locating hole of branched pipe inserted base mould bottom surface, then first order outer mold is enclosed within the first order step of base mould, to pour into mould from the perfusing hole of first order outer mold containing hepatocellular matrix solution, thrombin solution is adopted to be cross-linked after filling first order outer mold and base mould gap, removal first order outer mold and first order inner mold after completing, form stable ground floor cellular matrix layer, wherein containing the blood vessel access formed by first order Ink vessel transfusing mould and the neural inner mold of the first order and neural channel,
5) matrix solution containing Schwann cell is poured into into neural channel;
6) respectively total arm of the neural inner mold of second level Ink vessel transfusing mould and the second level is arranged on the outer mold of the second level, branched pipe inserts blood vessel access on the ground floor cellular matrix layer that has been shaped and neural channel, cover second level outer mold, to pour into mould from the perfusing hole of second level outer mold containing hepatocellular matrix solution, thrombin solution is adopted to be cross-linked after filling second level outer mold and ground floor cellular matrix lamellar spacing, second level outer mold is removed successively after being cross-linked, the neural inner mold in the second level and second level Ink vessel transfusing mould, form the stable second layer cellular matrix layer with second level blood vessel access and second level neural channel,
7) step 5 is repeated) and step 6), obtain third layer cellular matrix layer, the 4th confluent monolayer cells hypothallus and layer 5 cellular matrix layer respectively;
8) synthesis macromolecular solution is sprayed onto five confluent monolayer cells substrate layer surface formed thereby, after extraction, forms synthesized polymer material layer, remove base mould, namely form the liver precursor of band Pyatyi multiple-limb passage.
Embodiment 2: preparation band level Four multiple-limb blood vessel and neural cardiac precursor
1) base mould and level Four outer mold prepared containing level Four bench floor projection is combined by three-dimensional printing technology and Machining Technology, dies cavity is class heart surface, material therefor is politef, composite fibre materials is used to prepare level Four inner mold respectively, first order inner mold contains 1 branched pipe, and all the other inner molds all contain 4 branched pipes;
2) respectively sodium alginate and calcium chloride are dissolved in PBS solution and make 3% sodium alginate soln as matrix solution and 2% calcium chloride solution, buy endotheliocyte, Schwann cell and myocardial cell and make cell suspending liquid, it is 2 × 10 that the cell density of endotheliocyte suspension, Schwann cell suspension is
7individual/ml, the cell density of myocardial cell suspension is 1.2 × 10
7individual/ml, matrix solution and cell suspending liquid are mixed and made into celliferous matrix solution by 5:5 volume ratio respectively, be equipped with concentration be the different glycol solution of 30% polyester/tetra-of 30% (w/v) as synthesis macromolecular solution, and add the paclitaxel of 3% (w/v);
3) carry out vascularization process at part inner mold and make Ink vessel transfusing mould, specific practice applies the matrix solution of one deck containing endotheliocyte at mold surface, be cross-linked with calcium chloride solution, spray endotheliocyte cell suspending liquid subsequently and form endothelial layer, then synthesize macromolecular solution in endothelial layer surface sprinkling and extract, so far form the hypothallus-endothelial layer-synthesis macromolecule layer three-decker containing endotheliocyte at mold surface, all the other inner molds do not process is used as neural inner mold;
4) total arm of first order Ink vessel transfusing mould and the neural inner mold of the first order is arranged on first order outer mold, in the locating hole of branched pipe inserted base mould bottom surface, then first order outer mold is enclosed within the first order step of base mould, matrix solution containing myocardial cell is poured into mould from the perfusing hole of first order outer mold, calcium chloride solution is adopted to be cross-linked after filling first order outer mold and base mould gap, the neural inner mold of removal first order outer mold and the first order and first order Ink vessel transfusing mould after completing, form the stable ground floor cellular matrix layer containing first order blood vessel access and first order neural channel,
5) matrix solution containing Schwann cell is poured into into first order neural channel;
6) respectively total arm of the neural inner mold of second level Ink vessel transfusing mould and the second level is arranged on the outer mold of the second level, branched pipe inserts blood vessel access on the ground floor cellular matrix layer that has been shaped and neural channel, cover second level outer mold, matrix solution containing myocardial cell is poured into mould from the perfusing hole of second level outer mold, calcium chloride solution is adopted to be cross-linked after filling second level outer mold and ground floor cellular matrix lamellar spacing, the neural inner mold of second level outer mold and the second level and second level Ink vessel transfusing mould is removed successively after being cross-linked, form the second layer cellular matrix layer stablized with second level blood vessel access and second level neural channel,
7) step 5 is repeated) and step 6), obtain third layer cellular matrix layer and the 4th confluent monolayer cells hypothallus respectively;
8) synthesis macromolecular solution is sprayed onto four confluent monolayer cells substrate layer surface formed thereby, after extraction, forms synthesized polymer material layer, namely form half cardiac precursor;
9) repeat above-mentioned steps and prepare other half cardiac precursor again, two and half cardiac precursor are combined by synthesis macromolecular solution infiltration adhesion, namely forms the liver precursor with the band level Four multiple-limb passage of cavity structure.
Embodiment 3: the breast precursor preparing band ten grades of multiple-limb blood vessels
1) base mould and ten grades of outer molds of ten grades of steps are contained with three-dimensional printing technology preparation, dies cavity is class breast curved surface, material therefor is Merlon, polyurethane material is used to prepare ten grades of inner molds respectively, first order inner mold contains 1 branched pipe, and all the other inner molds all contain 1 or 2 branched pipes;
2) 2g gelatin and the mixing of 1g Fibrinogen are dissolved in 20mL DMEM culture fluid and make gelatin/fibrinogen solution as matrix solution, 10IU/mL thrombin solution; Purchase endotheliocyte, fat stem cell make cell suspending liquid, and cell density is all 1 × 10
7individual/ml; Matrix solution and cell suspending liquid are mixed and made into celliferous matrix solution by 4:6 volume ratio respectively; Outfit concentration is that the polylactic acid/aqueous isopropanol of 30% (w/v) synthesizes macromolecular solution as synthesis, and adds the sodium citrate of 30% (w/v);
3) vascularization process is carried out to part inner mold and make Ink vessel transfusing mould, concrete grammar applies the matrix solution of one deck containing endotheliocyte at mold surface, be cross-linked with thrombin solution, spray endotheliocyte cell suspending liquid subsequently and form endothelial layer, then synthesize macromolecular solution in endothelial layer surface sprinkling synthesis and extract, so far form the hypothallus-endothelial layer-synthesis synthesis macromolecule layer three-decker containing endotheliocyte at mold surface, all the other inner molds do not process;
4) total arm of first order inner mold is arranged on first order outer mold, in the locating hole of branched pipe inserted base mould bottom surface, then first order outer mold is enclosed within the first order step of base mould, the matrix solution of fatty stem cell is poured into mould from the perfusing hole of first order outer mold, thrombin solution is adopted to be cross-linked after filling first order outer mold and base mould gap, the neural inner mold of removal first order outer mold and the first order and first order Ink vessel transfusing mould after completing, form the stable ground floor cellular matrix layer containing first order blood vessel access and first order Common passageway,
5) respectively total arm of second level inner mold is arranged on the outer mold of the second level, branched pipe inserts blood vessel access on the ground floor cellular matrix layer that has been shaped and Common passageway, cover second level outer mold, the matrix solution of fatty stem cell is poured into mould from the perfusing hole of second level outer mold, thrombin solution is adopted to be cross-linked after filling second level outer mold and ground floor cellular matrix lamellar spacing, second level outer mold and inner mold is removed successively after being cross-linked, form the second layer cellular matrix layer stablized with second level blood vessel access and second level Common passageway,
7) step 5 is repeated) and step 6), obtain third layer respectively to the tenth confluent monolayer cells hypothallus;
8) synthesis synthesis macromolecular solution is sprayed onto ten confluent monolayer cells substrate layer surface formed thereby, synthesis synthesized polymer material layer is formed after extraction, the histoorgan be made up of multi-layer cellular hypothallus is taken out from bed die, namely form the breast precursor of band ten grades of branch vessel and Common passageway, Common passageway can directly as lactiferous ducts road.
Embodiment 4: the lung precursor preparing band six grades of multiple-limb blood vessels, nerve and tracheas
1) base mould and six grades of outer molds of six grades of steps are contained with three-dimensional printing technology preparation, dies cavity is class brain flexure face, material therefor is polyethylene, polytetrafluoroethylmaterial material is used to prepare six grades of inner molds respectively, first order inner mold contains 1 branched pipe, and all the other inner molds all contain 2 branched pipes;
2) 0.5g collagen is dissolved in 10mL normal saline and makes collagen solution as matrix solution; Preparation mass body volume concentrations is the glutaraldehyde solution of 1%; Buy endotheliocyte, Schwann cell and pneumonocyte and make cell suspending liquid, cell density is all 5 × 106/ml; Matrix solution and cell suspending liquid are mixed and made into celliferous matrix solution by 2:8 volume ratio respectively; Be equipped with the polyurethane/ethylene glycol solution of 5% as synthesis synthesis macromolecular solution, and add the paclitaxel of 5%;
3) vascularization process is carried out to part inner mold and make Ink vessel transfusing mould, specific practice applies the matrix solution of one deck containing endotheliocyte at mold surface, be cross-linked with glutaraldehyde solution, spray endotheliocyte cell suspending liquid subsequently and form endothelial layer, then synthesize macromolecular solution in endothelial layer surface sprinkling and extract, so far form the hypothallus-endothelial layer-synthesis macromolecule layer three-decker containing endotheliocyte at mold surface, all the other inner molds do not process respectively as neural inner mold and trachea inner mold;
4) total arm of first order inner mold is arranged on first order outer mold, in the locating hole of branched pipe inserted base mould bottom surface, then first order outer mold is enclosed within the first order step of base mould, matrix solution containing pneumonocyte is poured into mould from the perfusing hole of first order outer mold, glutaraldehyde solution is adopted to be cross-linked after filling first order outer mold and base mould gap, removal first order outer mold after completing, the neural inner mold of the first order, first order tracheal strips mould and first order Ink vessel transfusing mould, formed stable containing first order blood vessel access, the ground floor cellular matrix layer of air pipe passway and neural channel,
5) matrix solution containing Schwann cell is poured into into first order neural channel;
6) respectively total arm of the neural inner mold of second level Ink vessel transfusing mould and the second level is arranged on the outer mold of the second level, branched pipe inserts blood vessel access on the ground floor cellular matrix layer that has been shaped and neural channel, cover second level outer mold, matrix solution containing pneumonocyte is poured into mould from the perfusing hole of second level outer mold, glutaraldehyde solution is adopted to be cross-linked after filling second level outer mold and ground floor cellular matrix lamellar spacing, the neural inner mold of second level outer mold and the second level is removed successively after being cross-linked, second level Ink vessel transfusing mould and second level organ inner mold, formed stable with second level blood vessel access, the second layer cellular matrix layer of neural channel and air pipe passway,
7) step 5 is repeated) and step 6), obtain third layer respectively to layer 6 cellular matrix layer;
8) synthesis macromolecular solution is sprayed onto six confluent monolayer cells substrate layer surface formed thereby, synthesized polymer material layer is formed after extraction, the histoorgan be made up of multi-layer cellular hypothallus is taken out from bed die, namely forms the lung precursor of band six grades of multiple-limb blood vessels, nerve and tracheas;
Embodiment 5: the bladder precursor preparing band three grades of multiple-limb blood vessels, nerve and urethras
1) base mould and three grades of outer molds of three grades of steps are contained with three-dimensional printing technology preparation, dies cavity is class kidney curved surface, material therefor is politef, nylon is used to prepare three grades of Ink vessel transfusing moulds and neural inner mold respectively, first order inner mold contains 1 branched pipe, and all the other two-stage inner molds all contain 5 branched pipes;
2) preparation contains the mixed solution of 5% Fibrinogen and 4% sodium alginate as matrix solution, 10IU/mL thrombin solution and 1% calcium chloride solution; Buy endotheliocyte, Schwann cell and bladder cells and make cell suspending liquid, cell density is 2 × 107/ml; Matrix solution and cell suspending liquid are mixed and made into celliferous matrix solution by 6:4 volume ratio respectively; Be equipped with 30% polyurethane/TEG solution as synthesis macromolecular solution, and add the paclitaxel of 5%;
3) vascularization process is carried out to part inner mold and make Ink vessel transfusing mould, specific practice applies the matrix solution of one deck containing endotheliocyte at mold surface, be cross-linked respectively with thrombin solution and calcium chloride solution, spray endotheliocyte cell suspending liquid subsequently and form endothelial layer, then synthesize macromolecular solution in endothelial layer surface sprinkling and extract, so far form the hypothallus-endothelial layer-synthesis macromolecule layer three-decker containing endotheliocyte at mold surface, all the other inner molds do not process respectively as neural inner mold and urethra inner mold;
4) total arm of first order inner mold is arranged on first order outer mold, in the locating hole of branched pipe inserted base mould bottom surface, then first order outer mold is enclosed within the first order step of base mould, matrix solution containing bladder cells is poured into mould from the perfusing hole of first order outer mold, thrombin solution and calcium chloride solution is adopted to be cross-linked respectively after filling first order outer mold and base mould gap, removal first order outer mold and first order inner mold after completing, formed stable containing first order blood vessel access, the ground floor cellular matrix layer of neural channel and urethra,
5) matrix solution containing Schwann cell is poured into into first order neural channel;
6) respectively by second level Ink vessel transfusing mould, total arm of the neural inner mold of second level urethra inner mold and the second level is arranged on the outer mold of the second level, branched pipe inserts the blood vessel access on the ground floor cellular matrix layer that has been shaped, urethra and neural channel, cover second level outer mold, matrix solution containing bladder cells is poured into mould from the perfusing hole of second level outer mold, thrombin solution and calcium chloride solution is adopted to be cross-linked respectively after filling second level outer mold and ground floor cellular matrix lamellar spacing, second level outer mold and second level inner mold is removed successively after being cross-linked, formed stable with second level blood vessel access, the second layer cellular matrix layer of neural channel and urethra,
7) step 5 is repeated) and step 6), obtain third layer cellular matrix layer;
8) synthesis macromolecular solution is sprayed onto three confluent monolayer cells substrate layer surface formed thereby, synthesized polymer material layer is formed after extraction, the histoorgan be made up of multi-layer cellular hypothallus is taken out from bed die, namely forms the bladder precursor of band three grades of multiple-limb blood vessels, nerve and urethras.
Claims (8)
1. prepare a particular manufacturing craft for the histoorgan precursor of band multiple-limb passage, it is characterized in that: described particular manufacturing craft comprises the outer mold (101) of base mould (102), at least one-level and the inner mold (103) of at least one-level; Described base mould (102) inside is for becoming large stepped from bottom to up successively, and step number is identical with outer mold progression, and base mould bottom surface is plane, concave surface or convex surface, and is distributed with locating hole; Every grade of outer mold is equipped with installing hole and perfusing hole (402), the ladder of the external dimensions of outer mold and the corresponding progression of base mould to surround the size in face identical, the inner chamber of next stage outer mold and the inner chamber of upper level outer mold seamlessly transit, and the inner chamber of outer mold has the profile identical with presetting histoorgan; Every grade of inner mold comprises at least one branch-like structure, and its number is identical with the number of the installing hole of respective stages outer mold, each branch-like structure comprises total arm (301) and branched pipe (302), total arm of each branch-like structure and the installing hole of respective stages outer mold form matching relationship, the number of the branched pipe of most higher level's inner mold is 1, and total manifold size of next stage inner mold is identical with the branched pipe size of adjacent upper level inner mold.
2., according to a kind of particular manufacturing craft preparing the histoorgan precursor of band multiple-limb passage according to claim 1, it is characterized in that: the centrage of the perfusing hole of described multistage outer mold is positioned at same straight line.
3., according to a kind of particular manufacturing craft preparing the histoorgan precursor of band multiple-limb passage according to claim 1, it is characterized in that: the material of described base mould and outer mold is hard synthesized polymer material or metal material; Inner mold is the solid tubulose be made up of inanimate object toxic materials, and the material of this solid tubulose is synthetic fibers, polyethylene, Merlon, polrvinyl chloride or polyurethane.
4. adopt a method for the histoorgan precursor of particular manufacturing craft as claimed in claim 1 preparation band multiple-limb passage, it is characterized in that the method comprises the steps:
1) preparation quality volumetric concentration be the hydrogel solution of 1% ~ 30% as matrix solution, extract or buy zooblast make cell suspending liquid, cell suspending liquid density is 1 × 10
6~ 5 × 10
7individual/ml, is mixed and made into celliferous matrix solution in 1 ~ 9:9 ~ 1 by volume by matrix solution and cell suspending liquid, is dissolved in by synthesized polymer material in organic solvent and makes the synthesis macromolecular solution that mass percentage concentration is 1% ~ 30%;
2) three-dimensional printing technology is utilized to prepare particular manufacturing craft, total arm of first order inner mold is arranged on the installing hole of first order outer mold, first order outer mold is enclosed within the first order step of base mould, the branched pipe of first order inner mold is arranged in the locating hole of base mould bottom surface, celliferous matrix solution is poured into mould from the perfusing hole of first order outer mold, physics or chemical crosslinking or polymerization is adopted after filling first order outer mold and base mould gap, celliferous matrix solution in mould is cross-linked, first order outer mold and first order inner mold is removed successively after being cross-linked, form stable ground floor cellular matrix layer, wherein containing the first order branched bottom formed by first order inner mold,
3) total arm of second level inner mold is arranged on the installing hole of second level outer mold, second level outer mold is enclosed within the second level step of base mould, the branched pipe of second level inner mold aims at the passage on the ground floor cellular matrix layer that has been shaped, celliferous matrix solution is poured into mould from the perfusing hole of second level outer mold, physics or chemical crosslinking or polymerization is adopted after filling second level outer mold and ground floor cellular matrix lamellar spacing, celliferous matrix solution is cross-linked, second level outer mold and second level inner mold is removed successively after being cross-linked, form the stable second layer cellular matrix layer with second level passage,
4) step 3 is repeated), form third layer respectively to the cellular matrix layer of the superiors, obtain the multi-layer cellular hypothallus of the mutually through band multiple-limb passage of inter-layer passages;
5) synthesis macromolecular solution is sprayed onto multi-layer cellular substrate layer surface formed thereby, after extraction, forms synthesis macromolecule layer, remove base mould, namely form the histoorgan precursor of band multiple-limb passage.
5. according to the preparation method of the histoorgan precursor of band multiple-limb passage according to claim 4, it is characterized in that: the multiple-limb passage in the histoorgan precursor of described band multiple-limb passage forms blood vessel, nerve, bile duct, trachea or urinary catheter in histoorgan precursor incubation.
6. according to the preparation method of the histoorgan precursor of band multiple-limb passage according to claim 4, it is characterized in that: described hydrogel solution is one or more in gelatin, Fibrinogen, collagen, chitosan, sodium alginate, hyaluronic acid and fibronectin solution.
7. according to the preparation method of the histoorgan precursor of band multiple-limb passage according to claim 4, it is characterized in that: described synthesized polymer material adopts the complex of one or more materials in polyurethane, polycaprolactone, Merlon, polylactic acid, Polyethylene Glycol, polylactic acid, polyester, polyhydroxy acid ester and lactic acid and ethanol copolymer, organic solvent for dissolving synthesized polymer material adopts TEG, ethylene glycol, isopropyl alcohol or Isosorbide-5-Nitrae-dioxane.
8. according to the preparation method of the histoorgan precursor of band multiple-limb passage according to claim 4, it is characterized in that: in described celliferous matrix solution and synthesis macromolecular solution, add one or more in endothelial cell growth factor (ECGF), cell transfer factor or hepatocyte growth factor, and one or more in heparin, paclitaxel or sulfated chitosan.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510419730.XA CN105012050A (en) | 2015-07-16 | 2015-07-16 | Method and special mould for preparing tissue and organ precursor with multi-branch channels |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510419730.XA CN105012050A (en) | 2015-07-16 | 2015-07-16 | Method and special mould for preparing tissue and organ precursor with multi-branch channels |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105012050A true CN105012050A (en) | 2015-11-04 |
Family
ID=54402613
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510419730.XA Pending CN105012050A (en) | 2015-07-16 | 2015-07-16 | Method and special mould for preparing tissue and organ precursor with multi-branch channels |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105012050A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107286359A (en) * | 2017-06-27 | 2017-10-24 | 上普博源（北京）生物科技有限公司 | A kind of hydrogel of heterogenous multilayer structure and preparation method thereof |
| CN107446818A (en) * | 2016-09-14 | 2017-12-08 | 四川蓝光英诺生物科技股份有限公司 | Printing equipment, Method of printing and the lumen organization's construct of lumen organization's construct |
| CN110327134A (en) * | 2019-07-26 | 2019-10-15 | 中国医科大学 | Detachable particular manufacturing craft and the method for preparing multiple-limb channel complex organ precursor |
| CN111921016A (en) * | 2020-07-20 | 2020-11-13 | 中国医科大学 | Artificial organ manufacturing mold and artificial organ manufacturing method |
| CN113229994A (en) * | 2021-05-10 | 2021-08-10 | 中国医科大学 | Special combined die for preparing complex organ and method for preparing complex organ |
| CN113229993A (en) * | 2021-05-10 | 2021-08-10 | 中国医科大学 | Detachable combined die and method for preparing complex organ with multi-branch channel |
| CN113528338A (en) * | 2021-07-19 | 2021-10-22 | 中国医科大学 | Special combined die for drug screening and use method thereof |
| CN113528337A (en) * | 2021-07-19 | 2021-10-22 | 中国医科大学 | Combined die for organ manufacturing and drug screening and use method thereof |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008140413A1 (en) * | 2007-05-15 | 2008-11-20 | NEUROGEN MEDICAL INNOVATION I UMEå | Fibrin-based nerve repair conduit and method of producing the same |
| CN101632841A (en) * | 2009-08-21 | 2010-01-27 | 暨南大学 | Tissue engineering scaffold containing alginate, glycosaminoglycan and collagen and preparation method thereof |
| WO2011011962A1 (en) * | 2009-07-31 | 2011-02-03 | 清华大学 | Method for preparing complex multi-layer tissue organ precursor |
| CN102512261A (en) * | 2011-12-28 | 2012-06-27 | 清华大学 | Preparation method for complex organ precursors on basis of combination molds |
| CN102631709A (en) * | 2012-04-13 | 2012-08-15 | 清华大学 | Method for preparing complex organ precursor with branch vessel network |
| US20120209308A1 (en) * | 2006-03-31 | 2012-08-16 | Spiration, Inc. | Articulable anchor |
| CN102871771A (en) * | 2012-09-04 | 2013-01-16 | 清华大学 | Method for preparing fusiform complicated organ precursor by using rotary assembling die |
| CN102895702A (en) * | 2012-11-09 | 2013-01-30 | 四川大学华西医院 | Composite manual bile duct and preparation method thereof |
| CN103654999A (en) * | 2013-12-04 | 2014-03-26 | 上海纳米技术及应用国家工程研究中心有限公司 | Neutral restoration catheter support of multilayer structure and manufacturing method of neutral restoration catheter support |
| CN104207859A (en) * | 2014-09-16 | 2014-12-17 | 清华大学 | Method and special equipment utilizing spin accumulation method to prepare tissues and organs |
-
2015
- 2015-07-16 CN CN201510419730.XA patent/CN105012050A/en active Pending
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120209308A1 (en) * | 2006-03-31 | 2012-08-16 | Spiration, Inc. | Articulable anchor |
| WO2008140413A1 (en) * | 2007-05-15 | 2008-11-20 | NEUROGEN MEDICAL INNOVATION I UMEå | Fibrin-based nerve repair conduit and method of producing the same |
| WO2011011962A1 (en) * | 2009-07-31 | 2011-02-03 | 清华大学 | Method for preparing complex multi-layer tissue organ precursor |
| CN101632841A (en) * | 2009-08-21 | 2010-01-27 | 暨南大学 | Tissue engineering scaffold containing alginate, glycosaminoglycan and collagen and preparation method thereof |
| CN102512261A (en) * | 2011-12-28 | 2012-06-27 | 清华大学 | Preparation method for complex organ precursors on basis of combination molds |
| CN102631709A (en) * | 2012-04-13 | 2012-08-15 | 清华大学 | Method for preparing complex organ precursor with branch vessel network |
| CN102871771A (en) * | 2012-09-04 | 2013-01-16 | 清华大学 | Method for preparing fusiform complicated organ precursor by using rotary assembling die |
| CN102895702A (en) * | 2012-11-09 | 2013-01-30 | 四川大学华西医院 | Composite manual bile duct and preparation method thereof |
| CN103654999A (en) * | 2013-12-04 | 2014-03-26 | 上海纳米技术及应用国家工程研究中心有限公司 | Neutral restoration catheter support of multilayer structure and manufacturing method of neutral restoration catheter support |
| CN104207859A (en) * | 2014-09-16 | 2014-12-17 | 清华大学 | Method and special equipment utilizing spin accumulation method to prepare tissues and organs |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107446818A (en) * | 2016-09-14 | 2017-12-08 | 四川蓝光英诺生物科技股份有限公司 | Printing equipment, Method of printing and the lumen organization's construct of lumen organization's construct |
| CN107446818B (en) * | 2016-09-14 | 2021-05-07 | 四川蓝光英诺生物科技股份有限公司 | Lumen tissue construct printing device, lumen tissue construct printing method and lumen tissue construct |
| CN107286359A (en) * | 2017-06-27 | 2017-10-24 | 上普博源（北京）生物科技有限公司 | A kind of hydrogel of heterogenous multilayer structure and preparation method thereof |
| CN107286359B (en) * | 2017-06-27 | 2020-05-22 | 上普博源（北京）生物科技有限公司 | Hydrogel with heterogeneous multilayer structure and preparation method thereof |
| CN110327134A (en) * | 2019-07-26 | 2019-10-15 | 中国医科大学 | Detachable particular manufacturing craft and the method for preparing multiple-limb channel complex organ precursor |
| CN111921016A (en) * | 2020-07-20 | 2020-11-13 | 中国医科大学 | Artificial organ manufacturing mold and artificial organ manufacturing method |
| CN113229994A (en) * | 2021-05-10 | 2021-08-10 | 中国医科大学 | Special combined die for preparing complex organ and method for preparing complex organ |
| CN113229993A (en) * | 2021-05-10 | 2021-08-10 | 中国医科大学 | Detachable combined die and method for preparing complex organ with multi-branch channel |
| CN113528338A (en) * | 2021-07-19 | 2021-10-22 | 中国医科大学 | Special combined die for drug screening and use method thereof |
| CN113528337A (en) * | 2021-07-19 | 2021-10-22 | 中国医科大学 | Combined die for organ manufacturing and drug screening and use method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105012050A (en) | Method and special mould for preparing tissue and organ precursor with multi-branch channels | |
| Gaspar et al. | Advanced bottom‐up engineering of living architectures | |
| Zhao et al. | Review on the vascularization of organoids and organoids-on-a-C hip | |
| WO2016138701A1 (en) | Artificial blood vessel and preparation method thereof | |
| CN110327134A (en) | Detachable particular manufacturing craft and the method for preparing multiple-limb channel complex organ precursor | |
| US8663625B2 (en) | Diffusively permeable monolithic biomaterial with embedded microfluidic channels | |
| CN106178110B (en) | Ice glue three-dimensional structure, preparation method and application | |
| CN105985925B (en) | A kind of global function artificial organs fitting body and its preparation and cultural method | |
| CN104207859B (en) | Rotation method of piling is utilized to prepare method and the special equipment of histoorgan | |
| CN111481320B (en) | Method for preparing liver precursor by special combined die for preparing complex organ | |
| CN102631709B (en) | Method for preparing complex organ precursor with branch vessel network | |
| Ochoa et al. | An overview of the pathology and approaches to tissue engineering | |
| WO2011011962A1 (en) | Method for preparing complex multi-layer tissue organ precursor | |
| CN110177584A (en) | Low temperature gel 3D bracket and its production method | |
| CN111921016B (en) | Artificial organ manufacturing mold and artificial organ manufacturing method | |
| US11944719B2 (en) | Thin film interposition of basement membrane scaffolds | |
| CN106031800A (en) | Hollow stent for inducing regeneration and moulding of cartilage/bone tissues | |
| Wu et al. | Creation of a vascular system for organ manufacturing | |
| CN101492655A (en) | Vascularized fat depot based on partition and construction method thereof | |
| CN104658395A (en) | Heart simulation structure as well as forming method and special mold thereof | |
| CN106361463A (en) | Liver tissue simulation structure and forming method thereof | |
| Li et al. | Collagen-based bioinks for regenerative medicine: Fabrication, application and prospective | |
| US9629939B2 (en) | Collagenous foam materials | |
| CN211243906U (en) | Detachable special mould | |
| CN102871771B (en) | Method for preparing fusiform complicated organ precursor by using rotary assembling die |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20151104 |
|
| WD01 | Invention patent application deemed withdrawn after publication |