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Biological gel membrane for cell culture as well as preparation method and application thereof

A biological gel and cell culture technology, applied in the field of cell culture, can solve the problems of high price, dangerous clinical use, complicated operation, etc., and achieve the effect of reducing the amount of enzyme used, reducing cell loss, and improving amplification efficiency

Inactive Publication Date: 2013-05-08
黑龙江省重生生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is expensive and complicated to operate. More importantly, it still needs to be digested with enzymes when collecting cells. The enzyme digestion process is likely to adversely affect the stability of stem cell karyotypes. At the same time, cells are used as seeds for cell therapy or tissue engineering. When the cells are used, the incorporation of animal-derived digestive enzyme components may cause potential danger to their clinical use

Method used

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  • Biological gel membrane for cell culture as well as preparation method and application thereof
  • Biological gel membrane for cell culture as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Preparation of a biogel membrane for cell culture

[0033] (1) Weigh 2g of chlorinated chitosan and dissolve it in 98ml of deionized water to prepare a chlorinated chitosan solution with a mass fraction of 2%, and stir until it is completely dissolved (for later use).

[0034] (2) Weigh 2g of gelatin and dissolve it in 98ml of deionized water to prepare a gelatin solution with a mass fraction of 2%, stir until it is completely dissolved (for later use).

[0035] (3) A certain volume of Matrigel solution (purchased from BD Company, item number: 354234), to which a certain amount of deionized water was added to form a Matrigel solution with a protein concentration of 9 mg / ml. The entire operation process should be performed on ice.

[0036] (4) Measure 10ml of the chitosan chloride solution prepared in step (1) and 10ml of the gelatin solution prepared in step (2), mix and stir, filter and sterilize for later use.

[0037] (5) Dissolve 1 g of sodium glyceropho...

Embodiment 2

[0039] Embodiment 2: the preparation of a kind of biogel film for cell culture

[0040] (1) Weigh 2g of chlorinated chitosan and dissolve it in 98ml of deionized water to prepare a chlorinated chitosan solution with a mass concentration of 2%, and stir until it is completely dissolved (for later use).

[0041] (2) Weigh 2g of a certain amount of gelatin and dissolve it in 98ml of deionized water to prepare a gelatin solution with a mass concentration of 2%, and stir until it is completely dissolved (for later use).

[0042] (3) Take a certain volume of homemade Matrigel solution (prepared according to the method in Example 5), and add a certain amount of deionized water to it to form a Matrigel solution with a protein concentration of 9 mg / ml. The entire operation process needs to be performed on ice.

[0043] (4) Measure 10ml of the chitosan chloride solution prepared in step (1) and 5ml of the gelatin solution prepared in step (2), mix and stir, filter and sterilize for late...

Embodiment 3

[0046] Embodiment 3: A kind of preparation for the biogel membrane of cell culture

[0047] (1) Weigh 2g of chlorinated chitosan and dissolve it in 98ml of deionized water to prepare a chlorinated chitosan solution with a mass concentration of 2%, and stir until it is completely dissolved (for later use).

[0048] (2) Weigh 2g of a certain amount of gelatin and dissolve it in 98ml of deionized water to prepare a gelatin solution with a mass concentration of 2%, and stir until it is completely dissolved (for later use).

[0049] (3) Take a certain volume of self-made Matrigel solution (prepared according to the method in Example 5), and add a certain amount of deionized water to it to form a Matrigel solution with a concentration of 9 mg / ml. The entire operation process needs to be performed on ice.

[0050] (4) Measure 10ml of the chitosan chloride solution prepared in step (1) and 10ml of the gelatin solution prepared in step (2), mix and stir, filter and sterilize for later ...

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Abstract

The invention discloses a biological gel membrane for cell culture as well as a preparation method and an application thereof. The biological gel membrane contains biocompatible Matrigel, chitosan chloride and gelatin. The biological gel membrane can simulate structure, composition, physical properties and functions of an in-vivo cell basilar membrane, and can support proliferation of multiple cells. The biological gel membrane is liquid at room temperature, becomes a gel membrane when the temperature is more than 34 DEG C and is semi-liquid when the temperature is reduced, so cells can be recovered by changing the temperature, the consumption of enzyme is reduced, loss and functional change of the cells are reduced, and the amplification efficiency of the cells is improved. The invention also provides the method for preparing the biological gel membrane and the application in the cell culture.

Description

technical field [0001] The invention relates to a biogel film for cell culture, a preparation method and application thereof, and belongs to the technical field of cell culture. Background technique [0002] Cell expansion is a necessary prerequisite for the development of cell therapy and tissue engineering. In recent years, a variety of new measures for large-scale culture of adherent cells have been widely used in large-scale culture of engineered cell lines, that is, by simulating cells in vivo The basic starting point for culturing cells is to control their expansion and directional differentiation. At present, materials with good biocompatibility are used to simulate the extracellular matrix and promote the expansion of cells in vitro, mainly including hyaluronic acid, collagen, chitosan, alginate, gelatin, etc. In recent years, Matrigel has received more and more attention in cell culture due to its unique properties. Matrigel is isolated from EHS mouse tumors and ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08L89/00C08L5/08C08K5/521C08J3/075C12N5/071C12N5/0735
Inventor 李萌
Owner 黑龙江省重生生物科技有限公司
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