Method for constructing high-throughput cell mass model through droplet microfluidic gravitational positioning

A cell cluster and microfluidic technology, applied in the field of flux-based micro-cell cluster construction, can solve the problems of limited application, risk of aseptic operation, lack of consistency and flux, etc., to improve efficiency and facilitate recovery Effect

Pending Publication Date: 2022-03-15
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional macroscopic cell cluster construction methods usually lack consistency and throughput, which in turn limits their application in drug and toxicity screening
The microarray chip partially solves the problems of consistency and throughput, but the operation involving the recovery of the cell mass during the construction of the cell mass on it usually needs to be performed manually, and may include operations in which a large area is exposed to air, which poses risks to aseptic operations
In addition, cell clusters in arrays usually need to be cultured in solution in microwells for recovery, which limits the use of extracellular matrix-like matrices in micro-organ construction.

Method used

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  • Method for constructing high-throughput cell mass model through droplet microfluidic gravitational positioning
  • Method for constructing high-throughput cell mass model through droplet microfluidic gravitational positioning
  • Method for constructing high-throughput cell mass model through droplet microfluidic gravitational positioning

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Embodiment 1

[0035] In this embodiment, a method for constructing a high-throughput cell cluster model by liquid droplet microfluidic gravity positioning includes the following steps:

[0036] a. Microfluidic preparation of cell microdroplets:

[0037] Prepare the mixed solution of gel and cells as the aqueous phase, where the gel is natural gel, synthetic gel, composite gel, including all of which have reversible temperature-sensitive phase transition behavior in physiological solutions and are below the gel-sol transition temperature Form a physical gel, be in a liquid state above the gel-sol transition temperature, and be able to maintain cell activity; prepare oil containing surfactants as the oil phase; the mass concentration of the gel in the aqueous phase mixed solution is 3% -20% at a cell concentration of 1 x 10 5 -1×10 8 cells / ml, uniform cell-containing micro-droplets are produced by the droplet microfluidic chip;

[0038] b. Synchronized cell gravity sedimentation:

[0039]...

Embodiment 2

[0046] This embodiment is basically the same as Embodiment 1, especially in that:

[0047] A high-throughput method for constructing cell clusters, using the above-mentioned microfluidic chip to generate gel balls to wrap cells, specifically includes the following steps:

[0048] Such as figure 1 As shown, microfluidics is used to form uniform cell microdroplets. The gels here are natural gels, synthetic gels, and composite gels, including all of which have reversible temperature-sensitive phase transition behavior in physiological solutions, and in gel - A gel that forms a physical gel below the sol transition temperature, is in a liquid state above the gel-sol transition temperature, and is able to maintain cell viability. Cells in the exponential growth phase were first digested and resuspended with fresh culture medium to adjust the cell concentration to 1×10 8 cells / mL, use the culture medium with adjusted cell concentration to prepare 10% methacrylic anhydride gelatin ...

Embodiment 3

[0053] This embodiment is basically the same as the above-mentioned embodiment, and the special features are:

[0054] A high-throughput method for constructing cell clusters, using the above-mentioned microfluidic chip to generate gel balls to wrap cells, specifically includes the following steps:

[0055] Such as figure 1 As shown, microfluidics is used to form uniform cell microdroplets. The gels here are natural gels, synthetic gels, and composite gels, including all of which have reversible temperature-sensitive phase transition behaviors in physiological solutions, and in gels - A gel that forms a physical gel below the sol transition temperature, is in a liquid state above the gel-sol transition temperature, and is able to maintain cell viability. Cells in exponential growth phase were first digested and resuspended with fresh culture medium to adjust the cell concentration to 5×10 6 cells / mL, use the culture medium with adjusted cell concentration to configure 10% me...

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Abstract

The invention discloses a method for constructing a high-throughput cell mass model through droplet microfluidic gravitational positioning, multi-cell micro-droplets are prepared in a high-throughput manner by using a droplet microfluidic method, and the size of a cell mass is regulated by adjusting the cell concentration. And standing in a cell culture box, settling by virtue of gravity to form a cell accumulation body, and adhering the cultured cells to form a cell cluster. And the time for primarily forming the cell mass is only 12 hours. The gel is good in biocompatibility, physical gel is formed below the gel-sol transition temperature, and the physical gel is heated to the gel-sol transition temperature and is in a liquid state. Cell mass culture is carried out below the gel-sol transformation temperature, and cell mass recovery is carried out above the gel-sol transformation temperature. The cell cluster constructed by the method has the advantages of high flux, short time consumption, good consistency and simplicity and convenience in recovery, and pollution caused by contact with air is reduced in a closed chip environment. The construction method of the cell mass model can be really applied to the fields of drug activity, substance toxicity screening and the like.

Description

technical field [0001] The invention relates to a method for constructing a biological model, in particular to a method for constructing flux-quantified miniature cell clusters that can be used in high-throughput drug and material safety tests. Background technique [0002] The biological model is the basic model necessary for substance toxicity and drug effectiveness testing. Traditionally used biological models are mainly animal models and artificially cultured cell models. There are significant biological differences between animal models and humans, so the test results of animal models are difficult to directly apply to humans, and even completely different experimental results can be obtained, making it difficult to replace human experiments. Artificially cultured cell models are closer to the human body than animals, but artificial culture is usually carried out in a two-dimensional state, while many cells in the human body grow in a three-dimensional and dynamic envi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/36C12N5/00
CPCC12M23/16C12M41/00C12N5/00
Inventor 徐碧漪刘畅唐涛李文张阿方
Owner SHANGHAI UNIV
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