CN117467157A - Porous hydrogel microcarrier for 3D cell culture and preparation method and application thereof - Google Patents

Porous hydrogel microcarrier for 3D cell culture and preparation method and application thereof Download PDF

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CN117467157A
CN117467157A CN202311414311.8A CN202311414311A CN117467157A CN 117467157 A CN117467157 A CN 117467157A CN 202311414311 A CN202311414311 A CN 202311414311A CN 117467157 A CN117467157 A CN 117467157A
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porous hydrogel
cell culture
hydrogel microcarrier
microcarrier
cell
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李智立
周德重
雍海洋
张雨荷
郭蕊
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Xian Jiaotong University
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Abstract

The invention discloses a porous hydrogel microcarrier for 3D cell culture and a preparation method and application thereof, belonging to the field of biomedical materials. The porous hydrogel microcarrier disclosed by the invention is formed by crosslinking a cyclic polymer with a natural biological material. The method comprises the steps of synthesizing a cyclic polymer, preparing the polymer through controllable active polymerization of vinyl monomers, controlling chemical composition and structure, and preparing the hydrogel microcarrier with natural biological materials through simple crosslinking reaction under the microfluidic technology. Experiments prove that the porous hydrogel microcarrier has good biocompatibility and mechanical property, pore diameter intercommunication can realize cell adhesion and growth, can effectively realize free exchange of substances such as nutrient substances and wastes, and can be used in the fields of cell culture, cell expansion, cell encapsulation, cell transplantation, regenerative medicine and the like.

Description

Porous hydrogel microcarrier for 3D cell culture and preparation method and application thereof
Technical Field
The invention belongs to the field of biomedical materials, relates to a porous hydrogel microcarrier and a preparation method thereof, and in particular relates to a porous hydrogel microcarrier for 3D cell culture and a preparation method and application thereof.
Background
Human mesenchymal stem cells are a class of multipotent cells with proliferation, differentiation potential and ability to replicate self-renewal. The use of stem cell therapy to provide new cell therapies for a number of currently untreatable diseases, human disease modeling or drug discovery has great application prospects, such as blood system diseases, nervous system diseases, cardiovascular diseases, diabetes, bone joint diseases, and the like. In recent years, the clinical application of stem cell therapy is rapidly increasing, and statistical studies indicate that tens of millions to billions of stem cells are required per kilogram of body weight of each patient, and methods based on cell therapy and tissue regeneration for these diseases are required to produce high quality and large amounts of stem cells, and induce differentiation to meet clinical demands. Traditional two-dimensional (2D) culture techniques are simple and standardized to operate, but due to their limitations, they fail to provide clinically desirable cell doses and are prone to abnormal cell division, loss of phenotype, etc., ultimately resulting in their lost differentiation potential.
Hydrogels, because of their unique three-dimensional (3D) network structure, versatility (porosity, permeability, mechanical properties), can mimic the regulation of cell proliferation and differentiation similar to the in vivo extracellular matrix Environment (ECM), have been successfully used for stem cell culture and expansion. Over the past few decades, a wide variety of hydrogel-based 3D Microcarriers (MCs) have been developed, providing higher specific surface areas for stem cell expansion to meet the rapidly growing clinical needs.
However, as a stem cell adhesion culture medium, the structure, physicochemical properties, and surface microenvironment of the material may affect cell adhesion, growth, proliferation, and differentiation behavior. Most of the microcarriers on the market at present have complex process, single function and can not be modified, such as microstructure, mechanical property and the like. Therefore, it is critical to design a suitable microcarrier, choosing the optimal materials, the simplest process and the large scale amplification technology to meet the expectations of quality, safety, efficacy and commercial viability.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a porous hydrogel microcarrier for 3D cell culture, and a preparation method and application thereof, so as to solve the technical problems that the microcarrier in the prior art is complex in process, single in function and incapable of being modified.
In order to achieve the above purpose, the invention is realized by adopting the following technical scheme:
the invention discloses a porous hydrogel microcarrier for 3D cell culture, which is prepared by crosslinking a cyclic polymer synthesized by natural biological materials and vinyl monomers; wherein, the mass ratio of the natural biological material to the cyclic polymer synthesized by vinyl monomer is 1: (0.2-5).
Preferably, the vinyl monomer is polyethylene glycol diacrylate, diethylene glycol dimethacrylate, tetraethylene glycol diacrylate, butylene glycol diacrylate, pentaerythritol triacrylate, pentaerythritol tetraacrylate, methoxypolyethylene glycol acrylate, hydroxypropyl methacrylate, trimethylolpropane triacrylate, glycidyl methacrylate, methyl acrylate, isocyanoethyl methacrylate, 2- (dimethylamino) ethyl acrylate, 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) ethyl ester, hydroxyethyl acrylate, disulfide acrylate, 2-acrylic acid-1, 10-decanediyl ester, acrylamide, N-dimethyl bisacrylamide or N, N' -bis (propionyl) cystamine.
Preferably, the vinyl monomer has an average molecular weight of 100 to 2000Da.
Preferably, the natural biological material comprises one or more of gelatin, gelatin derivatives, hyaluronic acid, collagen, glycoproteins, laminin, fibronectin, alginate and alginate derivatives.
The invention also discloses a preparation method of the porous hydrogel microcarrier for 3D cell culture, which comprises the following steps: the method comprises the steps of taking a cyclic polymer synthesized by a natural biological material and vinyl monomers as a disperse phase by utilizing a microfluidic technology, forming W/O emulsion with a continuous phase, regulating the flow rate ratio of the disperse phase to the continuous phase, emulsifying to form uniform liquid drops, collecting and solidifying the uniform liquid drops for at least more than 24 hours to fully crosslink the uniform liquid drops, preparing micron-sized spherical particles, and preparing the porous hydrogel microcarrier for 3D cell culture by a pore-forming process.
Preferably, the dispersed phase is a cyclic polymer synthesized by vinyl monomers in an aqueous phase solution, or a natural biological material in an aqueous phase solution, or a cyclic polymer synthesized by vinyl monomers and a natural biological material in an aqueous phase solution.
Preferably, the continuous phase is an organic phase solution comprising an organic solvent and a surfactant.
Preferably, the crosslinking reaction is a chemical crosslinking method, a physical crosslinking method, an enzyme-catalyzed crosslinking method, or a photocrosslinking method.
Preferably, the pore-forming process employs a solvent evaporation method, a calcium carbonate template method, a phase separation method, a freeze-drying method, or a gas foaming method.
The invention also discloses application of the porous hydrogel microcarrier for 3D cell culture, which comprises any one of the following steps:
1) For culturing and expanding stem cells;
2) For encapsulation of drug molecules or cells;
3) Used for cell transplantation.
Compared with the prior art, the invention has the following beneficial effects:
the porous hydrogel microcarrier for 3D cell culture is formed by crosslinking a cyclic polymer synthesized by vinyl monomers with a natural biological material, and the polymer is prepared by controllable active polymerization of the vinyl monomers and has controllable chemical composition and structure. On one hand, the raw material components are all raw materials commonly used in the biomedical field, so that the prepared porous hydrogel microcarrier has good biocompatibility. Experiments prove that the porous hydrogel microcarrier has good biocompatibility and mechanical property, pore diameter intercommunication can realize cell adhesion and growth, can effectively realize free exchange of substances such as nutrient substances and wastes, and can be used in the fields of cell culture, cell expansion, cell encapsulation, cell transplantation, regenerative medicine and the like. On the other hand, since the terminal of the cyclic polymer synthesized by the vinyl monomer contains unreacted double bonds, the terminal can be modified and modified by a plurality of methods such as Michael addition, click chemistry and the like, the function of the polymer can be further improved, and the adjustable porous hydrogel microcarrier can be prepared, so that the application field of the adjustable porous hydrogel microcarrier is expanded. Therefore, the porous hydrogel microcarrier can effectively solve the technical problems that the prior art is complex in process, single in function and incapable of being modified.
Drawings
FIG. 1 is a schematic diagram of cyclic polyethylene glycol synthesis;
FIG. 2a is the polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 1: chain transfer agent (CPADB) of 200: gel permeation chromatogram of cyclic polyethylene glycol in the reaction process;
FIG. 2b is the polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 2: chain transfer agent (CPADB) is 100: gel permeation chromatogram of cyclic polyethylene glycol in the reaction process;
FIG. 3a is the polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 1: chain transfer agent (CPADB) of 200:1, nuclear magnetic spectrum of the cyclic polyethylene glycol;
FIG. 3b is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 2: chain transfer agent (CPADB) is 100:1, nuclear magnetic spectrum of the cyclic polyethylene glycol;
FIG. 4 is a schematic diagram of a microfluidic device;
FIG. 5a is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 1: chain transfer agent (CPADB) of 200:1, optical microscopy pictures of micron-sized spherical particles formed by the synthesized cyclic polymer and thiolated gelatin;
FIG. 5b is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 2: chain transfer agent (CPADB) is 100:1, optical microscopy pictures of micron-sized spherical particles formed by the synthesized cyclic polymer and thiolated gelatin;
FIG. 6a is the polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 1: chain transfer agent (CPADB) of 200:1, electron microscope pictures of porous hydrogel microcarriers formed by synthesized cyclic polymers and thiolated gelatin;
FIG. 6b is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 2: chain transfer agent (CPADB) is 100:1, electron microscope pictures of porous hydrogel microcarriers formed by synthesized cyclic polymers and thiolated gelatin;
FIG. 7a is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 1: chain transfer agent (CPADB) of 200:1, culturing iMSCs by using a porous hydrogel microcarrier formed by the synthesized cyclic polymer and thiolated gelatin;
FIG. 7b is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 2: chain transfer agent (CPADB) is 100:1, culturing iMSCs by using a porous hydrogel microcarrier formed by the synthesized cyclic polymer and thiolated gelatin;
FIG. 8a is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 1: chain transfer agent (CPADB) of 200:1, culturing the cell activity diagram of the iMSCs by using a porous hydrogel microcarrier formed by the synthesized cyclic polymer and thiolated gelatin;
FIG. 8b is polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) of example 2: chain transfer agent (CPADB) is 100: cell activity profile of the synthetic cyclic polymer and thiolated gelatin-formed porous hydrogel microcarriers in culture of iMSCs at 1.
Detailed Description
In order that those skilled in the art will better understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, shall fall within the scope of the present invention.
It should be noted that the terms "first," "second," and the like in the description and the claims of the present invention and the above figures are used for distinguishing between similar objects and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used may be interchanged where appropriate such that the embodiments of the invention described herein may be implemented in sequences other than those illustrated or otherwise described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the attached drawing figures:
the invention discloses a porous hydrogel microcarrier, which consists of a cyclic polymer and a natural biological material, and is a hydrogel system.
The invention discloses a preparation method of a porous hydrogel microcarrier, which comprises the following steps:
1) A certain amount of vinyl monomer and a chain transfer agent 4-cyano-4- (thiobenzoyl) valeric acid (CPADB) are added into a three-mouth bottle filled with a reaction solvent butanone, and the monomer is fully dissolved through magnetic stirring, wherein the reaction charging mole ratio of the monomer and the CPADB is (50-500): 1, a step of;
the vinyl monomer is at least one of the following: polyethylene glycol diacrylate (average molecular weight 100-2000 Da), diethylene glycol dimethacrylate, tetraethylene glycol diacrylate, butylene glycol diacrylate, pentaerythritol triacrylate, pentaerythritol tetraacrylate, methoxypolyethylene glycol acrylate, hydroxypropyl methacrylate, trimethylolpropane triacrylate, glycidyl methacrylate, methyl acrylate, isocyanoethyl methacrylate, 2- (dimethylamino) ethyl acrylate, 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) ethyl ester, hydroxyethyl acrylate, disulfide acrylate, 2-acrylic acid-1, 10-decanediyl ester, acrylamide, N, N-dimethyl bisacrylamide, N, N' -bis (propionyl) cystamine.
2) Deoxidizing for 30min by using a nitrogen bubbling method;
3) Under the condition of introducing nitrogen, an initiator azo diisobutyl cyanide (AIBN) is rapidly added, and the nitrogen is continuously introduced for 5min; wherein, the feeding mole ratio of AIBN to CPADB is 0.5:1.
4) Immersing the three-mouth bottle into an oil bath preheated to 70 ℃ to start reaction;
5) Monitoring polymer molecular weight using gel permeation chromatography;
6) Stopping the reaction when the reaction molecular weight is close to 10000-100000 g/mol, and quenching free radicals;
7) The product was purified by precipitation to give a cyclic polymer.
8) Mixing the annular polyethylene glycol with the natural biological material at normal temperature or respectively placing the mixture into a microfluidic device to obtain a disperse phase;
the natural biological material is at least one of the following: gelatin, gelatin derivatives, hyaluronic acid, collagen, glycoproteins, laminin, fibronectin, alginate and alginate derivatives;
wherein, the reaction of the cyclic polyethylene glycol and the natural biological material is fed (0.2-5): 1.
9) Referring to fig. 4, by injecting different phases (a dispersed phase and a continuous phase) into a microfluidic chip at a controlled flow rate using a syringe pump, a water-in-oil (W/O) emulsion is formed in the microchip, and a flow rate ratio of the dispersed phase to the continuous phase is adjusted, and uniform droplets are formed by emulsification of the continuous phase;
the pore-forming process is at least one of the following: solvent evaporation, calcium carbonate template, phase separation, freeze drying, and gas foaming;
10 Collecting and solidifying the liquid drops for at least more than 24 hours to enable the liquid drops to be fully crosslinked to form micron-sized spherical particles;
the crosslinking reaction is at least one of the following: chemical crosslinking, physical crosslinking, enzymatic crosslinking.
11 The process also comprises the steps of washing the micron-sized spherical particles for a plurality of times after the reaction is finished, and drying or freeze-drying in air.
The specific implementation is as follows:
example 1
1) 10mmol of polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) and 0.05mmol of chain transfer agent 4-cyano-4- (thiobenzoyl) pentanoic acid (CPADB) are added into a three-necked flask filled with 50mL of reaction solvent butanone and the monomers are fully dissolved by magnetic stirring, deoxygenated for 30 minutes by using a nitrogen bubbling method, then 0.025mmol of initiator Azobisisobutyronitrile (AIBN) is rapidly added under nitrogen bubbling conditions, and nitrogen is continuously introduced for 5 minutes; reacting for 9h at 70 ℃; the reaction solution was precipitated by adding to diethyl ether and n-hexane solvent, filtered, and freeze-dried, and the molecular weight of the finally obtained product was 24000g/mol. Wherein, the reaction charging ratio of polyethylene glycol diacrylate (PEGDA, molecular weight is 700 g/mol), chain transfer agent (CPADB) and initiator (AIBN) is 200:1:0.5.
referring to FIG. 1, a schematic synthesis of the cyclic polyethylene glycol is shown.
The chemical structural formula of the cyclic polyethylene glycol prepared by the preparation method is as follows:
where p=35, n=13, m=15.
2) Dissolving 20mg of the obtained annular polyethylene glycol and 50mg of thiolated gelatin in 1mL of Phosphate Buffer Solution (PBS) respectively to prepare a disperse phase (aqueous phase), mixing the two disperse phases or respectively placing the two disperse phases in a microfluidic device, injecting different phases (the disperse phase and the continuous phase) into the microfluidic chip at a controlled flow rate by using an injection pump, forming W/O emulsion in the microchip, adjusting the flow rate ratio of the disperse phase to the continuous phase, forming uniform liquid drops by emulsifying the continuous phase, collecting and solidifying the liquid drops for at least more than 24 hours, and fully crosslinking the liquid drops to form micron-sized spherical particles. And after the reaction is finished, washing the micron-sized spherical particles for multiple times, and drying or freeze-drying in air to obtain the porous hydrogel microcarrier.
Example 2
1) 3mmol of polyethylene glycol diacrylate (PEGDA, molecular weight 700 g/mol) and 0.03mmol of chain transfer agent 4-cyano-4- (thiobenzoyl) pentanoic acid (CPADB) are added into a three-necked flask filled with 50mL of reaction solvent butanone and the monomers are fully dissolved by magnetic stirring, deoxygenated for 30 minutes by using a nitrogen bubbling method, then 0.015mmol of initiator Azobisisobutyronitrile (AIBN) is rapidly added under nitrogen bubbling conditions, and nitrogen bubbling is continued for 5 minutes; reacting for 10 hours at 70 ℃; the reaction solution was precipitated by adding it to diethyl ether and n-hexane solvent, filtered, and freeze-dried to obtain a final product having a molecular weight of 29000g/mol. Wherein, the reaction charging ratio of polyethylene glycol diacrylate (PEGDA, molecular weight is 700 g/mol), chain transfer agent (CPADB) and initiator (AIBN) is 100:1:0.5.
referring to FIG. 1, a schematic synthesis of the cyclic polyethylene glycol is shown.
The chemical structural formula of the cyclic polyethylene glycol prepared by the preparation method is as follows:
where p=41, n= 9,m =31.
2) Dissolving 20mg of the obtained annular polyethylene glycol and 50mg of thiolated gelatin in 1mL of Phosphate Buffer Solution (PBS) respectively to prepare a disperse phase (aqueous phase), mixing the two disperse phases or respectively placing the two disperse phases in a microfluidic device, injecting different phases (the disperse phase and the continuous phase) into the microfluidic chip at a controlled flow rate by using an injection pump, forming W/O emulsion in the microchip, adjusting the flow rate ratio of the disperse phase to the continuous phase, forming uniform liquid drops by emulsifying the continuous phase, collecting and solidifying the liquid drops for at least more than 24 hours, and fully crosslinking the liquid drops to form micron-sized spherical particles. And after the reaction is finished, washing the micron-sized spherical particles for multiple times, and drying or freeze-drying in air to obtain the porous hydrogel microcarrier.
The invention is described in further detail below with reference to the attached drawing figures:
the porous hydrogel microcarrier disclosed by the invention has the following material properties and application:
1. the cyclic polyethylene glycol disclosed by the invention has good controllability and adjustability, and the molecular weight and the cyclizing degree of the obtained polymer are different by changing the ratio of polyethylene glycol diacrylate to chain transfer agent to initiator, so that the cyclic polyethylene glycol with different compositions and cyclizing degrees is obtained.
Referring to fig. 2a and 2b, there is shown gel permeation chromatography of the cyclic polyethylene glycol prepared according to the present invention;
referring to fig. 3a and 3b, the nuclear magnetic spectrum of the cyclic polyethylene glycol prepared by the invention is shown.
2. The porous hydrogel microcarrier disclosed by the invention has adjustable chemical composition and mechanical property, and is used for obtaining porous hydrogel microcarriers with different structures and properties by changing the molecular weight and cyclizing degree of the cyclic polymer or changing the crosslinking ratio of the cyclic polymer and thiolated gelatin.
Referring to fig. 5a and 5b, optical microscopy images of microcarriers made by cross-linking a cyclic polymer with thiolated gelatin according to the present invention are shown. It can be seen that micron-sized spherical particles with uniform size are formed by the microfluidic device, and the size is 100-300 μm;
referring to fig. 6a and 6b, electron microscopy images of microcarriers made by cross-linking a cyclic polymer with thiolated gelatin according to the present invention are shown. It can be seen that the microcarrier formed has a larger pore size and good connectivity, and the pore size varies from 3 to 8 μm.
In addition, the porous hydrogel microcarrier disclosed by the invention adopts PEGDA (molecular weight is 700 g/mol) as a reaction monomer, adopts CPADB as a chain transfer agent and AIBN as an initiator to form cyclic polyethylene glycol, and is formed by crosslinking with natural biological material thiolated gelatin, and the components are all raw materials commonly used in the biomedical field, so that the porous hydrogel microcarrier disclosed by the invention has good biocompatibility and can be used in the fields of medicine/protein purification, delivery, cell culture, cell expansion, cell encapsulation, cell transplantation, regenerative medicine and the like.
Thus, incubation of the porous hydrogel microcarriers at 37℃and 5% CO by placing the microcarriers in culture medium for at least 1h 2 Is then mixed with the iMSCs for culturing and expansion.
Referring to fig. 7a and 7b, fluorescence microscopy images of the microcarrier culture of iMSCs made by cross-linking the cyclic polymer with thiolated gelatin according to the present invention are shown. It can be seen that the microcarrier has good cell adhesion and biocompatibility.
Referring to FIGS. 8a and 8b, there are graphs showing the activity of the microcarrier culture iMSCs made by cross-linking a cyclic polymer with thiolated gelatin according to the present invention. It can be seen that both the iMSCs exhibited higher survival rates and amplification performance, exceeding 150% compared to the blank.
The above is only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited by this, and any modification made on the basis of the technical scheme according to the technical idea of the present invention falls within the protection scope of the claims of the present invention.

Claims (10)

1. A porous hydrogel microcarrier for 3D cell culture, characterized in that it is made by cross-linking a cyclic polymer synthesized from natural biological materials and vinyl monomers; wherein, the mass ratio of the natural biological material to the cyclic polymer synthesized by vinyl monomer is 1: (0.2-5).
2. The porous hydrogel microcarrier for 3D cell culture of claim 1, wherein the vinyl monomer is polyethylene glycol diacrylate, diethylene glycol dimethacrylate, tetraethylene glycol diacrylate, butylene glycol diacrylate, pentaerythritol triacrylate, pentaerythritol tetraacrylate, methoxypolyethylene glycol acrylate, hydroxypropyl methacrylate, trimethylolpropane triacrylate, glycidyl methacrylate, methyl acrylate, isocyanoethyl methacrylate, 2- (dimethylamino) ethyl acrylate, 2-methyl-2-acrylic acid-2- (2-methoxyethoxy) ethyl ester, hydroxyethyl acrylate, disulfides acrylate, 2-acrylic acid-1, 10-decanediyl ester, acrylamide, N-dimethyl bisacrylamide or N, N' -bis (propionyl) cystamine.
3. The porous hydrogel microcarrier for 3D cell culture of claim 1, wherein the vinyl monomer has an average molecular weight of 100-2000 Da.
4. The porous hydrogel microcarrier for 3D cell culture of claim 1, wherein the natural biological material comprises one or more of gelatin, gelatin derivatives, hyaluronic acid, collagen, glycoproteins, laminin, fibronectin, alginate, and alginate derivatives.
5. The method for preparing a porous hydrogel microcarrier for 3D cell culture according to any one of claims 1-4, comprising: the method comprises the steps of taking a cyclic polymer synthesized by a natural biological material and vinyl monomers as a disperse phase by utilizing a microfluidic technology, forming W/O emulsion with a continuous phase, regulating the flow rate ratio of the disperse phase to the continuous phase, emulsifying to form uniform liquid drops, collecting and solidifying the uniform liquid drops for at least more than 24 hours to fully crosslink the uniform liquid drops, preparing micron-sized spherical particles, and preparing the porous hydrogel microcarrier for 3D cell culture by a pore-forming process.
6. The method of claim 5, wherein the dispersed phase is a cyclic polymer synthesized from vinyl monomers in an aqueous solution, or a natural biomaterial in an aqueous solution, or a cyclic polymer synthesized from vinyl monomers and a natural biomaterial in an aqueous solution.
7. The method of claim 5, wherein the continuous phase is an organic phase solution comprising an organic solvent and a surfactant.
8. The method for preparing a porous hydrogel microcarrier for 3D cell culture according to claim 5, wherein the crosslinking reaction is chemical crosslinking, physical crosslinking, enzyme-catalyzed crosslinking or photo-crosslinking.
9. The method for preparing a porous hydrogel microcarrier for 3D cell culture of claim 5, wherein the pore-forming process employs solvent evaporation, calcium carbonate template, phase separation, lyophilization or gas foaming.
10. Use of a porous hydrogel microcarrier for 3D cell culture according to any one of claims 1-4, characterized in that it is any one of the following:
1) For culturing and expanding stem cells;
2) For encapsulation of drug molecules or cells;
3) Used for cell transplantation.
CN202311414311.8A 2023-10-27 2023-10-27 Porous hydrogel microcarrier for 3D cell culture and preparation method and application thereof Pending CN117467157A (en)

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