A method for bioreactor construction of a population of microdroplets
By constructing a microdroplet bioreactor and utilizing liquid-liquid phase separation and 3D printing technologies, the problems of poor mechanical performance and incompatibility of cell bioactivity in traditional immobilized cell reactors were solved, achieving stable cell culture and improved mechanical strength.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- XIANGSHENG TECH (NANJING) CO LTD
- Filing Date
- 2023-05-29
- Publication Date
- 2026-07-31
AI Technical Summary
Traditional immobilized cell reactors suffer from poor mechanical properties and incompatibility with cell bioactivity, which is particularly unfavorable for the culture of suspended cells, and also leads to cell leakage and reduced bioactivity.
A microdroplet swarm bioreactor construction method was adopted. Core-shell structured hydrogel microspheres loaded with cells were prepared by liquid-liquid phase separation technology and mixed with functional polymer materials to form core-shell droplets. The microdroplet swarm bioreactor was formed by 3D printing technology.
This improved the mechanical properties of the bioreactor, prevented cell leakage, maintained cell biological activity, and enabled 3D patterned cell culture.
Smart Images
Figure CN116656457B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of bioreactors, and more specifically, to a method for constructing a bioreactor using microdroplet clusters. Background Technology
[0002] A bioreactor is a device system that utilizes the biological functions of enzymes or organisms (such as microorganisms) to carry out biochemical reactions in vitro. It is a kind of biological function simulator, such as fermenters, immobilized enzymes, or immobilized cell reactors. Bioreactors are mainly used for plant and animal cell culture, microcarrier culture, and transgenic pharmaceutical engineering. Immobilized cell reactors mainly use the embedding method to immobilize cells, fixing cells within the pores of various gels, films, and polymer meshes. The embedding method for cell immobilization is relatively simple, has a high activity recovery rate, and the immobilized cell activity is maintained for a relatively long time. However, in traditional immobilized cell reactors, cell activity and mechanical strength are contradictory, especially unfavorable for the culture of suspension cells. Traditional immobilized cell reactors have poor mechanical properties, which is not conducive to preparation and molding, functional applications, and stable cell culture. Usually, directly increasing the polymer concentration, thereby increasing the cross-linking density and increasing the mechanical strength of the bioreactor, can lead to problems such as reduced biological activity of cells (especially suspension cells) embedded in hydrogels and cell leakage. Summary of the Invention
[0003] To address the aforementioned problems, this invention aims to resolve the incompatibility between the mechanical properties and cellular bioactivity of traditional hydrogel materials. For the first time, a method for constructing a microdroplet bioreactor is proposed. This novel bioreactor, through liquid-liquid phase separation, preserves the liquid cellular environment, while the core-shell structure and matrix enhance the material's mechanical properties and effectively prevent cellular leakage. It can be fabricated using 3D printing, enabling the 3D patterned culture of cells and other biological processes.
[0004] The technical solution of the present invention is: a method for constructing a microdroplet cluster bioreactor, the present invention comprising two components, the first component being a core-shell structured hydrogel microsphere loaded with cells, referred to as a cell carrier; the second component being a solution of a functional polymer material that can form a liquid-liquid phase separation with the solution inside the core, referred to as a matrix;
[0005] The construction method involves mixing the two components and transferring them into a template or using 3D printing or other methods to form and solidify the microdroplet clusters into a bioreactor.
[0006] Furthermore, the specific operation steps of the method for constructing a microdroplet swarm bioreactor are as follows:
[0007] (1) Using the principle of phase separation, droplet microfluidics, membrane emulsification and other methods are used to obtain the first component, namely the core-shell structured hydrogel microspheres used as cell carriers;
[0008] (2) Prepare the second component, namely the matrix;
[0009] It is mixed with a certain volume fraction of cell carrier to obtain a blend;
[0010] (3) The blend of the cell carrier and the matrix is transferred to a template or 3D printed to obtain the bioreactor of the microdroplet cluster.
[0011] Furthermore, in step (1), the specific preparation method of the core-shell structured hydrogel microspheres for cell carriers is as follows:
[0012] Using fluorinated oil as the oil phase, a functional polymer solution as the intermediate aqueous phase, and an aqueous polymer solution that can form a liquid-liquid phase separation with the intermediate aqueous phase as the inner aqueous phase, an oil-in-water-in-water (W / W / O) emulsion is generated in the channel of a microfluidic chip by utilizing the shearing action of the outer oil phase. This is achieved by a one-step method to generate a monodisperse dual emulsion that has been solidified.
[0013] Furthermore, the aqueous polymer solution in the inner aqueous phase can be one of sodium carboxymethyl cellulose, carboxymethyl cellulose, or dextran.
[0014] Furthermore, the functional polymer is one or more of the following: double-bonded gelatin, double-bonded Pluronic F-127, double-bonded PVA, and polyvinyl polyethylene glycol.
[0015] Furthermore, the polyvinyl polyethylene glycol is one or more of the following: multi-arm polyethylene glycol, dendritic polyethylene glycol ((mPEG)4-(PEG)2-MAL), and hyperbranched polyethylene glycol (hyperbranched polyethylene glycol diacrylate (HB-PEGDA)).
[0016] The multi-arm polyethylene glycol is one or more of the following: two-arm polyethylene glycol ((Propargyl-PEG)2-Allyl), three-arm polyethylene glycol (3Arm(PEG-Allyl3), four-arm polyethylene glycol (4Arm(PEG-Allyl)4), and six-arm polyethylene glycol (6Arm-PEG-DA)).
[0017] Furthermore, the droplet microfluidic can be one of a microtubular coaxial loop triple coaxial capillary chip, a PDMS chip, and a glass chip.
[0018] Furthermore, in step (2), the matrix is a functional polymer material that is printable and can form a liquid-liquid phase separation with the core-shell microdroplet core-phase polymer solution.
[0019] Furthermore, the matrix is a functional polymer material that is printable and can form a liquid-liquid phase separation with the core-shell structured hydrogel microspheres.
[0020] The matrix is one or more of double-bonded gelatin and double-bonded Pluronic F-127, and has the characteristic of being phase-separable from polymer solutions such as sodium carboxymethyl cellulose, carboxymethyl cellulose, and dextran.
[0021] Furthermore, the cell carrier and matrix blend can be used to adjust cell density and distribution by adjusting the ratio of cell carrier to matrix.
[0022] The ratio is 1:0.1 to 1.
[0023] Furthermore, the core-shell droplet biomaterial can independently encapsulate one or more types of cells, such as bacteria, algae, animal cells, and plant cells.
[0024] The beneficial effects of this invention are as follows: Firstly, the cell bio-disperses in an inner aqueous phase, and then separates the inner and outer aqueous phases through liquid-liquid phase separation to obtain a water-in-oil (W / W / O) emulsion. The outer aqueous phase is cross-linked using photocrosslinking, Michael addition, disulfide bond formation, Schiff base reaction, enzyme crosslinking, ionic crosslinking, or click chemistry to form core-shell droplets, encapsulating the cells inside the droplets. Then, a functional polymer material that forms a liquid-liquid phase separation with the solution inside the core is used as a matrix to fix the discrete core-shell droplets, avoiding the reduction in cell activity caused by compression during 3D printing and reducing cell escape. The method for preparing a core-shell droplet biomaterial described in this invention has advantages such as good biocompatibility, adjustable mechanical strength, and 3D cell culture. Attached image description:
[0025] Figure 1 This is a flowchart of the preparation process of the present invention;
[0026] Figure 2 This is a microscopic image of a core-shell droplet carrying an organism in an embodiment of the present invention;
[0027] Figure 3 This is a schematic diagram of microfluidic core-shell droplet preparation in an embodiment of the present invention;
[0028] Figure 4 This is a diagram showing the hydrodynamic properties of the core-shell droplet biomaterial in an embodiment of the present invention;
[0029] Among them, Figure (a) is the strain-modulus performance curve of the core-shell droplet biomaterial, Figure (b) is the angular velocity-modulus performance curve of the core-shell droplet biomaterial, and Figures (c) and (d) are the shear rate-viscosity performance curve and self-healing performance diagram of the core-shell droplet biomaterial before solidification.
[0030] Figure 5 This is a microscopic SEM image of the core-shell droplet biological living material in this invention. Detailed Implementation
[0031] To more clearly illustrate the technical solution of the present invention, the technical solution of the present invention will be further described in detail below with reference to the accompanying drawings:
[0032] Example 1
[0033] (1) Preparation of Bacillus subtilis core-shell droplets from double-bonded gelatin and sodium carboxymethyl cellulose (CMC):
[0034] Bacillus subtilis was activated in LB medium for 12 hours. 1 mL of the bacterial culture was centrifuged to remove the supernatant, and the solution was set aside. A 3% (w / v) surfactant (FE-surf) was dissolved in fluorinated oil and designated as the continuous phase P1 (oil phase). 80 mg of double-bonded gelatin and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL (20 mg / mL) of sodium chloride solution to prepare an 8% (w / v) double-bonded gelatin solution, designated as the dispersed phase P2. 10 mg of CMC was dissolved in 1 mL of water, and Bacillus subtilis was resuspended in this solution to prepare a 1% (w / v) solution. Let this be the dispersed phase P3. Connect the solution prepared above to the microfluidic chip through a PET tube. Adjust the flow rate of the oil phase P1 to 40 L / min, the flow rate of P3 to 4 μL / min, and the flow rate of P4 to 2 μL / min to prepare monodisperse droplets of Bacillus subtilis with a size of about 150 μm. Collect for 2 h and cure under 405 nm light for 5 min. Wash 5 times with 10% PFO fluorinated oil 7500 solution, then wash 5 times with pure fluorinated oil 7500, and finally wash twice with deionized water. Centrifuge to remove water to obtain core-shell droplets A1.
[0035] (2) Preparation of core-shell droplet living biological materials:
[0036] 160 mg of double-bonded gelatin and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 16% (w / v) double-bonded gelatin solution, denoted as matrix B1. Matrix B1 and core-shell droplet A1 were mixed evenly at 2 / 1 (v / v) at 35 degrees Celsius and then 3D printed at 4 degrees Celsius. After printing, the microdroplet bioreactor was obtained by curing under 405 nm light for 5 min.
[0037] Example 2
[0038] (1) Preparation of Bacillus subtilis core-shell droplets loaded with double-bonded gelatin and carboxymethyl cellulose (CM):
[0039] Bacillus subtilis was activated in LB medium for 12 hours. 1 mL of the bacterial culture was centrifuged to remove the supernatant, and the solution was set aside. A 3% (w / v) surfactant (FE-surf) was dissolved in fluorinated oil and designated as the continuous phase P1 (oil phase). 80 mg of double-bonded gelatin and 10 mg of blue light initiator (LAP) were dissolved in 1 mL (20 mg / mL) of sodium chloride solution to prepare an 8% (w / v) double-bonded gelatin solution, designated as the dispersed phase P2. 10 mg of CM was dissolved in 1 mL of water, and Bacillus subtilis was resuspended in this solution to prepare a 1% (w / v) dispersed phase P3. The solutions prepared above were then subjected to… PET tubing was connected to a microfluidic chip; the flow rates of P1 oil phase were adjusted to 40 L / min, P3 flow rates to 4 μL / min, and P3 flow rates to 2 μL / min to prepare monodisperse droplets of Bacillus subtilis with a size of about 150 μm. The droplets were collected for 2 h and cured by irradiation with 405 nm light for 5 min; the droplets were first washed 5 times with 10% PFO fluorinated oil 7500 solution, then washed 5 times with pure fluorinated oil 7500, and finally washed twice with deionized water. The water was removed by centrifugation to obtain core-shell droplets A2.
[0040] (2) Preparation of core-shell droplet living biological materials:
[0041] 160 mg of double-bonded gelatin and 10 mg of blue light initiator (LAP) were dissolved in 1 mL of water to prepare a 16% (w / v) double-bonded gelatin solution, denoted as matrix B2. Matrix B2 and core-shell droplet A2 were mixed evenly at 2 / 1 (v / v) at 35 degrees Celsius and then 3D printed at 4 degrees Celsius. After printing, the microdroplet bioreactor was obtained by curing under 405 nm light for 5 min.
[0042] Example 3
[0043] (1) Preparation of Bacillus subtilis core-shell droplets loaded with double-bonded gelatin and dextran:
[0044] Bacillus subtilis was activated in LB medium for 12 hours. 1 mL of the bacterial culture was centrifuged to remove the supernatant, and the solution was set aside. A 3% (w / v) surfactant (FE-surf) was dissolved in fluorinated oil and designated as the continuous phase P1 (oil phase). 80 mg of double-bonded gelatin and 10 mg of blue light initiator (LAP) were dissolved in 1 mL (20 mg / mL) of sodium chloride solution to prepare an 8% (w / v) double-bonded gelatin solution, designated as the dispersed phase P2. 40 mg of dextran was dissolved in 1 mL of water, and Bacillus subtilis was resuspended in this solution to prepare a 4% (w / v) dispersed phase P3. The solutions prepared above were then subjected to… PET tubing was connected to a microfluidic chip; the flow rates of P1 oil phase were adjusted to 40 L / min, P3 flow rates to 4 μL / min, and P4 flow rates to 2 μL / min to prepare monodisperse droplets of Bacillus subtilis with a size of about 150 μm. The droplets were collected for 2 h and cured by irradiation with 405 nm light for 5 min; the droplets were first washed 5 times with 10% PFO fluorinated oil 7500 solution, then washed 5 times with pure fluorinated oil 7500, and finally washed twice with deionized water. The water was removed by centrifugation to obtain core-shell droplets A3.
[0045] (2) Preparation of core-shell droplet living biological materials:
[0046] 160 mg of double-bonded gelatin and 10 mg of blue light initiator (LAP) were dissolved in 1 mL of water to prepare a 16% (w / v) double-bonded gelatin solution, denoted as matrix B3. Matrix B3 and core-shell droplet A3 were mixed evenly at 2 / 1 (v / v) at 35 degrees Celsius and then 3D printed at 4 degrees Celsius. After printing, the microdroplet bioreactor was obtained by curing under 405 nm light for 5 min.
[0047] Example 4
[0048] (1) Preparation of Bacillus subtilis core-shell droplets loaded with double bond Pluronic F-127 and dextran:
[0049] Bacillus subtilis was activated in LB medium for 12 hours. 1 mL of the bacterial culture was centrifuged to remove the supernatant, and the solution was set aside. A surfactant (FE-surf) was dissolved in fluorinated oil to prepare a 3% (w / w) continuous phase, designated as P1 (oil phase). 200 mg of double-bonded Pluronic acid was added... F-127 and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 20% (w / v) double-bonded Pluronic F-127 solution, denoted as dispersed phase P2. 40 mg of dextran was dissolved in 1 mL of water and used to resuspend Bacillus subtilis to prepare a 1% (w / v) dispersed phase, denoted as dispersed phase P3. The solutions prepared above were connected to a microfluidic chip via PET tubing. The flow rates of P1 (oil phase) were adjusted to 40 L / min, P3 (oil phase) to 4 μL / min, and P4 (oil phase) to 2 μL / min, to prepare monodisperse droplets of approximately 150 μm in size encapsulated with Bacillus subtilis. The droplets were collected for 2 h and cured under 405 nm light for 5 min. The core-shell droplets A4 were obtained by washing five times with 10% PFO fluorinated oil 7500 solution, then five times with pure fluorinated oil 7500, and finally twice with deionized water. Centrifugation was then used to remove water.
[0050] (2) Preparation of core-shell droplet living biological materials:
[0051] 400 mg of double-bonded Pluronic F-127 and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 40% (w / v) double-bonded gelatin solution, denoted as matrix B4. Matrix B4 and core-shell droplet A4 were mixed evenly at 2 / 1 (v / v) at 0 degrees Celsius and then 3D printed at 30 degrees Celsius. After printing, the microdroplet bioreactor was obtained by curing under 405 nm light for 5 min.
[0052] Example 5
[0053] (1) Preparation of Bacillus subtilis core-shell droplets loaded with double-bonded Pluronic F-127 and CMC:
[0054] Bacillus subtilis was activated in LB medium for 12 hours. 1 mL of the bacterial culture was centrifuged to remove the supernatant, and the solution was set aside. A surfactant (FE-surf) was dissolved in fluorinated oil to prepare a 3% (w / w) continuous phase, designated as P1 (oil phase). 200 mg of double-bonded Pluronic acid was added... F-127 and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 20% (w / v) double-bonded Pluronic F-127 solution, denoted as dispersed phase P2. 10 mg of CMC was dissolved in 1 mL of water and used to resuspend Bacillus subtilis to prepare a 1% (w / v) dispersed phase, denoted as dispersed phase P3. The solutions prepared above were connected to a microfluidic chip through PET tubing. The flow rates of P1 oil phase were adjusted to 40 L / min, P3 flow rates to 4 μL / min, and P3 flow rates to 2 μL / min to prepare monodisperse droplets of approximately 150 μm in size containing Bacillus subtilis. The droplets were collected for 2 h and cured by irradiation with 405 nm light for 5 min. The droplets were first washed 5 times with 10% PFO fluorinated oil 7500 solution, then washed 5 times with pure fluorinated oil 7500, and finally washed twice with deionized water. The water was removed by centrifugation to obtain core-shell droplets A5.
[0055] (2) Preparation of core-shell droplet living biological materials:
[0056] 400 mg of double-bonded Pluronic F-127 and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 40% (w / v) double-bonded gelatin solution, denoted as matrix B5. Matrix B5 and core-shell droplet A5 were mixed evenly at 2 / 1 (v / v) at 0 degrees Celsius and then 3D printed at 30 degrees Celsius. After printing, the microdroplet bioreactor was obtained by curing under 405 nm light for 5 min.
[0057] Example 6
[0058] (1) Preparation of Chlorella core-shell droplets loaded with double-bonded Pluronic F-127 and CMC:
[0059] Centrifuge 1 mL of algal solution to remove the supernatant and set aside. Dissolve the surfactant (FE-surf) in fluorinated oil to prepare a 3% (w / v) continuous phase, denoted as P1 (oil phase). Dissolve 200 mg of double-bonded Pluronic F-127 and 10 mg of blue photoinitiator (LAP) in 1 mL of water to prepare a 20% (w / v) double-bonded Pluronic F-127 solution, denoted as P2. Dissolve 10 mg of CMC in 1 mL of water and resuspend Chlorella in it to prepare a 1% (w / v) dispersed phase, denoted as P3. Connect the solutions prepared above to a microfluidic chip through PET tubing. Adjust the flow rate of P1 oil phase to 40 L / min, P3 flow rate to 4 μL / min, and P3 flow rate to 2 μL / min to prepare monodisperse droplets of Bacillus subtilis with a size of about 150 μm. Collect for 2 h and cure under 405 nm light for 5 min. First, wash five times with 10% PFO fluorinated oil 7500 solution, then wash five times with pure fluorinated oil 7500, and finally wash twice with deionized water. Centrifuge to remove water to obtain core-shell droplet A6.
[0060] (2) Preparation of core-shell droplet living biological materials:
[0061] 400 mg of double-bonded Pluronic F-127 and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 40% (w / v) double-bonded gelatin solution, denoted as matrix B6. Matrix B6 and core-shell droplet A6 were mixed evenly at 2 / 1 (v / v) at 0 degrees Celsius and then 3D printed at 30 degrees Celsius. After printing, the microdroplet bioreactor was obtained by curing under 405 nm light for 5 min.
[0062] Example 7
[0063] (1) Preparation of Bacillus subtilis & Chlorella core-shell droplets from double-bonded gelatin and sodium carboxymethyl cellulose (CMC):
[0064] Bacillus subtilis was activated in LB medium for 12 hours. 1 mL of the bacterial culture was centrifuged to remove the supernatant, and the solution was set aside. 1 mL of algal culture in the logarithmic growth phase was centrifuged to remove the supernatant, and the solution was set aside. A 3% (w / v) surfactant (FE-surf) was dissolved in fluorinated oil and designated as the continuous phase P1 (oil phase). 80 mg of double-bonded gelatin and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL (20 mg / mL) of sodium chloride solution to prepare an 8% (w / v) double-bonded gelatin solution, designated as the dispersed phase P2. 10 mg of CMC was dissolved in 1 mL of water, and Bacillus subtilis and Chlorella were resuspended in this solution, designated as the dispersed phase P3. The solution prepared above was connected to a microfluidic chip through a PET tube; the flow rates of the P1 oil phase were adjusted to 40 L / min, the P3 flow rate to 4 μL / min, and the P4 flow rate to 2 μL / min to prepare monodisperse droplets of Bacillus subtilis with a size of about 150 μm. The droplets were collected for 2 h and cured by irradiation with 405 nm light for 5 min; the droplets were first washed 5 times with 10% PFO fluorinated oil 7500 solution, then washed 5 times with pure fluorinated oil 7500, and finally washed twice with deionized water. The water was removed by centrifugation to obtain core-shell droplets A1.
[0065] (2) Preparation of core-shell droplet living biological materials:
[0066] 400 mg of double-bonded Pluronic F-127 and 10 mg of blue photoinitiator (LAP) were dissolved in 1 mL of water to prepare a 40% (w / v) double-bonded gelatin solution, denoted as matrix B6. Matrix B6 was mixed with core-shell droplets A6 at 2 / 1, 3 / 1, and 4 / 1 (v / v) ratios at 0°C until homogeneous. The mixture was then 3D printed at 30°C. After printing, the mixture was cured by irradiation with 405 nm light for 5 min to obtain bioreactors with microdroplet clusters of different cell distributions.
[0067] Finally, it should be understood that the embodiments described in this invention are only used to illustrate the principles of the embodiments of this invention; other variations may also fall within the scope of this invention; therefore, as examples rather than limitations, alternative configurations of the embodiments of this invention can be regarded as consistent with the teachings of this invention; correspondingly, the embodiments of this invention are not limited to the embodiments explicitly introduced and described in this invention.
Claims
1. A method for constructing a bioreactor using microdroplet swarms, characterized in that: The bioreactor comprises two components: the first component is a core-shell structured hydrogel microsphere loaded with cells, referred to as the cell carrier; the second component is a functional polymer material that is printable and can form a liquid-liquid phase separation with the core phase polymer solution of the core-shell structured hydrogel microsphere, referred to as the matrix. The specific preparation steps are as follows: Step (1): Using the principle of phase separation, the first component, namely the core-shell structured hydrogel microspheres used as cell carriers, is obtained by using droplet microfluidics. The specific preparation process of the hydrogel microspheres is as follows: using fluorinated oil of surfactant as oil phase, functional polymer solution as intermediate aqueous phase, and polymer aqueous solution that can form liquid-liquid phase separation with intermediate aqueous phase as inner aqueous phase, in the channel of microfluidic chip, the shearing action of the outer oil phase is used to generate water-in-oil (W / W / O) emulsion, and the monodisperse double emulsion is solidified in one step. Step (2): Prepare the second component, namely the matrix; mix the matrix and the cell carrier in proportion to obtain a blend containing the matrix and the cell carrier; Step (3): The prepared blend containing cell carrier and matrix is transferred to a template or 3D printed to form a bioreactor of microdroplet clusters after solidification.
2. The method for constructing a bioreactor using microdroplet swarms according to claim 1, characterized in that: The aqueous polymer solution in the internal aqueous phase mentioned in step (1) is one of carboxymethyl cellulose and dextran.
3. The method for constructing a bioreactor using microdroplet swarms according to claim 1, characterized in that: The functional polymer solution in the intermediate aqueous phase mentioned in step (1) is one or more of the following: double bond gelatin, double bond Pluronic F-127, double bond PVA, and polyvinyl polyethylene glycol.
4. The method for constructing a microdroplet swarm bioreactor according to claim 3, characterized in that: The polyvinyl polyethylene glycol is one or more of multi-arm polyethylene glycol, dendritic polyethylene glycol, or hyperbranched polyethylene glycol; The multi-arm polyethylene glycol is one or more of two-arm polyethylene glycol, three-arm polyethylene glycol, four-arm polyethylene glycol, and six-arm polyethylene glycol.
5. The method for constructing a microdroplet swarm bioreactor according to claim 1, characterized in that: In step (1), the droplet microfluidic is one of the following: microtubular coaxial loop triple coaxial capillary chip, PDMS chip, and glass chip.
6. The method for constructing a microdroplet swarm bioreactor according to claim 1, characterized in that: In step (2), the matrix is one of double-bonded gelatin and double-bonded Pluronic F-127.
7. The method for constructing a bioreactor using microdroplet swarms according to claim 1, characterized in that: The microdroplet bioreactor described in step (3) is used to independently encapsulate one or more cells from bacteria, algae, animal cells, and plant cells.