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Microfluidic device and method for preparing microgel by using microfluidic device

A microfluidic device and microgel technology, applied in the field of bioengineering, can solve problems affecting cell activity, product performance differences, and restrictions on wide application, and achieve strong stability

Active Publication Date: 2016-06-08
SHENZHEN HUA NOVA BIOTECH LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] 1) Photolithography technology, micro-template technology, liquid droplets based on centrifugal force: These traditional technologies are all based on batch production processes, which cannot achieve high-throughput continuous preparation, and there are differences in product performance between batches
[0011] 2) Oil-in-water (W / O) single emulsion droplet technology: This method cannot achieve continuous cell-loaded gel generation, but requires uninterrupted collection of products and breaking the emulsion to transfer cells to a biocompatible aqueous phase solution In the process, the process is time-consuming and labor-intensive
Also, prolonged exposure of cells to oils and surfactants can affect cell viability
[0012] 3) Water-in-water (W / W) emulsion technology: this system is limited to a specific combination of two immiscible aqueous solutes, such as dextran and polyethylene glycol, which limits the wide application of this method

Method used

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  • Microfluidic device and method for preparing microgel by using microfluidic device
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  • Microfluidic device and method for preparing microgel by using microfluidic device

Examples

Experimental program
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Effect test

Embodiment 1

[0074] The method of preparing monodisperse cell-carrying microgels using the ultra-thin shell-layer core-shell structure double emulsion as a template of the present invention will be further illustrated below with reference to the accompanying drawings.

[0075] Such as figure 1 Said, a microfluidic device comprising a first microchannel, a second microchannel, a third microchannel, an output channel, an ultraviolet light source and a collection container, the first microchannel, the second microchannel and the third microchannel The channels respectively have a sample sending end and an insertion end, and the insertion end of the second microchannel is inserted into the first microchannel without sealing from the sample sending end of the first microchannel to the insertion end; the insertion end of the third microchannel The second microchannel is unsealedly inserted into the second microchannel from the sample sending end to the insertion end to form a sleeve; one end of the ...

Embodiment 2

[0083] Using the microfluidic device described in Example 1, the method for preparing microgels includes the following steps:

[0084] (1) Configure internal phase, intermediate phase and external phase solutions:

[0085] The configuration of the internal phase solution: combine polyethylene glycol diacrylate (PEGDA) and light curing agent (1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2 under normal temperature and pressure -Methyl-1-propan-1-one) is dissolved in deionized water, the final concentration of PEGDA in the solution is 10w / v%, and the concentration of the light curing agent is 1w / v%.

[0086] Preparation of the mesophase solution: the mesophase is a mixed solution of oil and surfactant, and the Span80 surfactant solution is added to the mineral oil to make the concentration in the mineral oil 0.5v / v%.

[0087] Configuration of the external phase solution: Dissolve polyvinyl alcohol (PVA) in deionized water with a final mass concentration of 5%.

[0088] (2) Preparation of wate...

Embodiment 3

[0093] Using the microfluidic device described in Example 1, the method for preparing a microgel with living cells includes the following steps:

[0094] (1) Configure internal phase, intermediate phase and external phase solutions:

[0095] The configuration of the internal phase solution: methacrylate grafted gelatin and light curing agent (1-[4-(2-hydroxyethoxy)-phenyl]-2-hydroxy-2-methyl at room temperature and pressure -1-propan-1-one), dissolved in the cell culture medium to obtain the hydrogel prepolymer solution. The cell culture medium is Dulbecco's modified Eagle medium (DMEM, Sigma-Aldrich), plus 10% v / v fetal bovine serum (FBS, Gibco). Disperse the live cells cultured and expanded in vitro in the cell culture medium, and add the above-mentioned hydrogel prepolymer solution so that the cell concentration in the final solution is 1×10 6 Cells / ml, the concentration of methacrylate grafted gelatin is 10w / v%, and the concentration of light curing agent is 1w / v%.

[0096] Pre...

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Abstract

The invention provides a microfluidic device and a method for preparing microgel by using the microfluidic device. The method comprises the steps of (1) taking a solution containing live cells or bioactive molecules, a photocuring agent and a hydrogel prepolymer as an internal phase, taking a mixed solution containing oil and an surfactant as an intermediate phase, and taking a polyvinyl alcohol aqueous solution as an external phase; (2) respectively conveying the internal phase, the intermediate phase and the external phase into a corresponding microchannel of the microfluidic device through a micro pump or a micro injector to form a monodispersed water-in-oil-in-water double emulsion; (3) enabling the monodispersed water-in-oil-in-water double emulsion in the step (2) to pass through an output channel of the microfluidic device, and collecting the monodispersed water-in-oil-in-water double emulsion in the step (2) into a collection vessel filled with an aqueous solution, thus obtaining the microgel immobilizing with the live cells or the bioactive molecules. The preparation of the microgel immobilizing with cells by a one-step method is realized, the obtained microgel is controllable in size, and narrow in size distribution, and meanwhile the survival rate of cells is maintained.

Description

Technical field [0001] The invention belongs to the field of bioengineering, and relates to a microfluidic device and a method for preparing a microgel carrying living cells or biologically active factors by using the microfluidic device. Background technique [0002] For the repair of damaged or diseased human tissues or organs, the clinical tradition still relies on organ donation. However, the shortage of transplanted human tissues and the risk of causing infectious diseases make organ transplants face many problems. Taking the United States as an example, there are currently more than 74,000 patients waiting for organ transplantation each year, and only 21,000 people can get transplant operations each year. In recent years, the birth and development of regenerative medicine technology by means of tissue engineering and cell therapy has brought new hope to organ repair and reconstruction, and is expected to alleviate the actual problem of the shortage of organ donors. Among ...

Claims

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Application Information

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IPC IPC(8): A61L27/16A61L27/18A61L27/20A61L27/22A61L27/50A61L27/54C12M1/00
CPCA61L27/16A61L27/18A61L27/20A61L27/222A61L27/50A61L27/54A61L2400/06C12M1/00
Inventor 王华楠
Owner SHENZHEN HUA NOVA BIOTECH LTD
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