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Composite film based on egg white and methylacrylic acid derived polymer as well as application of composite film to cultivation of stem cells

A methacrylic acid and methacrylic acid ester technology, applied in animal cells, vertebrate cells, microorganisms, etc., can solve problems such as the lack of a clear and complete mechanism theory, achieve precise and controllable biological functions, and promote rapid proliferation. Effect

Active Publication Date: 2018-05-29
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The self-replication and differentiation behavior of stem cells is determined by many factors such as the state of the cell itself and the microenvironment in which the cells exist. So far, there is no clear and complete mechanism theory

Method used

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  • Composite film based on egg white and methylacrylic acid derived polymer as well as application of composite film to cultivation of stem cells
  • Composite film based on egg white and methylacrylic acid derived polymer as well as application of composite film to cultivation of stem cells
  • Composite film based on egg white and methylacrylic acid derived polymer as well as application of composite film to cultivation of stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Preparation and Cell Culture of Egg White-Methacrylated Gelatin Composite Film with Higher Degree

[0067] 1. A composite film based on egg white and gelatin with a higher degree of methacrylated, prepared by the following steps:

[0068] (1) Preparation of egg white solution: take fresh egg white, shake and mix for 7 min, centrifuge at 3000 r / min for 15 min, and then the layering is obvious, and take the middle clear solution for later use;

[0069] (2) Synthesis of GelMa material: Dissolve gelatin with a molecular weight of 20,000 in PBS buffer in a 50°C water bath to prepare a solution with a concentration of 15% (w / v), and slowly add methacrylic acid dropwise at a rate of 0.7 mL / min Anhydride, the ratio of which to gelatin is 7.5:1 (w / v), stirred and reacted in a water bath at 50°C for 12 h, then added 4 times the amount of PBS to stop the reaction. Put the obtained milky white liquid into a dialysis bag (8-14KD), and dialyze in distilled water for 7 days...

Embodiment 2

[0078] Example 2 Preparation and cell culture of egg white-low molecular weight PEGDA composite membrane

[0079] 1, a kind of composite film based on egg white and low molecular weight PEGDA is prepared by the following steps:

[0080] (1) Preparation of egg white solution: take a small hole on the eggshell, separate the egg white, oscillate and mix for 5 minutes, and then turn into a light yellow turbid shape. After centrifuging at 2000 r / min for 8 minutes, the layering is obvious, and the middle part of the solution is clear. That is the required egg white solution, separated for use;

[0081] (2) Synthesis of low-molecular-weight PEGDA: Mix PEG with a molecular weight of 200 and dichloromethane at a ratio of 10% (w / v), add triethylamine under ice-cooling, and use a constant-pressure funnel at a rate of 4 drops / 5 seconds. Add acryloyl chloride dropwise (the molar ratio of PEG: triethylamine: acryloyl chloride is 1:5:6); after the reaction, add 1 mol / L hydrochloric acid and...

Embodiment 3

[0087] Example 3 Preparation of egg white-methacrylated sugar (hyaluronic acid) composite film

[0088] 1. A composite film based on egg white and methacrylated sugars, prepared from the following steps:

[0089] (1) Preparation of egg white solution: take a small hole on the eggshell, separate the egg white, shake and mix for 10 minutes, and then it becomes light yellow and turbid. After centrifuging at 4000 r / min for 20 minutes, the layering is obvious, and the middle part of the solution is clear. That is the required egg white solution, separated for use;

[0090] (2) Synthesis of methacrylated hyaluronic acid material: dissolve hyaluronic acid with a molecular weight of 500,000 in PBS at 30°C in a water bath to prepare a saturated hyaluronic acid solution, and slowly drop it at a rate of 1 mL / min. Add methacrylic anhydride, the ratio of which to hyaluronic acid is 5:1 (w / v), stir and react at room temperature for 24 h, then add 4 times the amount of PBS to stop the react...

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Abstract

The invention discloses a composite film based on egg white and methylacrylic acid derived polymer as well as application of the composite film to cultivation of stem cells. The composite film comprises egg white and at least one methacrylic acid derivatization polymer. The egg white serves as a main raw material and reacts with the methacrylic acid derivatization polymer under the controlled condition to prepare the composite film, and the obtained composite film has high mechanical property and biological property and can serve as a scaffold material of the stem cells to support the growth of the stem cells; in addition, stable colloid is formed under the condition of low solid content and the strength of products is improved, so that the internal part of the composite film has sufficient space and the growth of the stem cells under different conditions is benefited; and besides, the performance of the scaffold material in various aspects can be further adjusted by adjusting the proportion of the raw materials, and the biological function can be controlled accurately. The product provided by the invention is reliable in performance, high in biocompatibility, simple and convenientin operation, high in repeatability and easy in industrialization.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and more specifically relates to a composite membrane based on egg white and methacrylic acid derivatized polymer, its preparation method, its application in culturing stem cells, and a method for cultivating stem cells on the membrane. Background technique [0002] Stem cells have attracted much attention in various fields such as life sciences, biology, and medicine because of their multi-directional differentiation potential. At present, research on stem cells is emerging in an endless stream around the world. From blood diseases to tissue damage repair to anti-aging, stem cell technology is gradually transitioning from clinical research to clinical application. New treatment options for major diseases such as liver disease. At the same time, as one of the key factors of tissue engineering, stem cells together with scaffold materials and growth factors constitute tissue engineering...

Claims

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Application Information

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IPC IPC(8): C08J5/18C08J3/28C08J9/28C08L89/00C08H1/00C08L71/02C08B5/08C08G65/332C08B37/08C12N5/0775
CPCC08B37/0072C08G65/3322C08H1/00C08J3/28C08J5/18C08J9/28C08J2201/0502C08J2371/02C08J2389/00C08J2405/08C08J2471/02C08J2489/00C12N5/0663C12N5/0664C12N2533/30
Inventor 吴钧黄钶晴欧阳宏顾志鹏
Owner SUN YAT SEN UNIV
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