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Preparation method of immobilized all-anthropogenic ECM coating matrix

A technology of coating and matrix, which is applied in the field of preparation of extracellular matrix, can solve problems such as insufficient effect, unsuitable for large-scale production, complex production process, etc., achieve improved lysis method, solve heterogeneity problem, and simple preparation method Effect

Pending Publication Date: 2015-06-24
JIANGYIN STEMEASY BIOTECH LTD
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  • Abstract
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  • Claims
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Problems solved by technology

But their production process is complex, not suitable for large-scale production, and the effect is not as good as feeder cells and Matrigel

Method used

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  • Preparation method of immobilized all-anthropogenic ECM coating matrix
  • Preparation method of immobilized all-anthropogenic ECM coating matrix
  • Preparation method of immobilized all-anthropogenic ECM coating matrix

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Example 1: Applying immobilized fully human ECM to human embryonic stem cell culture and result detection

[0041] The main instruments and models used in the experiment: ultrapure water instrument (SIMSV0000, Millipore); CO2 cell incubator (Thermo, 3111, the United States); ultra-clean bench (YJ-VS type, Wuxi Yijing Purification Equipment Co., Ltd.); fluorescence Inverted phase-contrast microscope (OLYMPUS, IX71, Japan); paraffin microtome (YD202A, Jinhua Yidi Medical Equipment Factory).

[0042] 1. Preparation of immobilized human ECM-coated matrix

[0043] 1.1 Obtain umbilical cord mesenchymal cells from human umbilical cord

[0044] 1) washing the fresh human umbilical cord obtained from a full-term human fetus with PBS three times;

[0045] 2) Cut the umbilical cord into small sections, and peel off two arteries and one vein;

[0046] 3) After removing the blood vessels, separate Wharton's jelly;

[0047] 4) Cut the isolated Wharton's jelly into small pieces,...

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Abstract

The invention relates to a preparation method of an immobilized all-anthropogenic ECM coating matrix, first, extracellular matrix can be obtained from human umbilical cord mesenchymal cells by cell lysis method, and finally the extracellular matrix is immobilized by methanol. The matrix is an immobilized extracellular matrix which is all-anthropogenic and ready-to-use, can be saved for a long time and is suitable for long-term culture growth of human embryonic stem cells, and can be used for in vitro amplification of clinical grade human embryonic stem cells.

Description

technical field [0001] The invention relates to a preparation method of immobilized extracellular matrix (extracellular matrix, ECM) derived from human umbilical cord. Background technique [0002] Human embryonic stem cells (hESC) are pluripotent stem cells with self-renewal ability. They are isolated from the inner cell mass of the blastocyst stage and have the ability to differentiate into three germ layers. Therefore, they are used for tissue repair and drug screening. Broad prospects. The culture system suitable for the clinical application of hESC must have the advantages of no animal origin, defined components, and suitable for large-scale production. However, hESCs are harsh on growth conditions. Only when the appropriate culture substrate and culture medium cooperate with each other can it maintain its undifferentiated state for a long time. In the traditional culture system, human embryonic stem cells are usually divided into feeder-layer and feeder-free culture...

Claims

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Application Information

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IPC IPC(8): C12N5/0735
Inventor 陈慧敏朱海林
Owner JIANGYIN STEMEASY BIOTECH LTD
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