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Mass culture method of umbilical cord mesenchymal stem cells

A kind of stem cell and culture method technology, which is applied in the field of mass culture of umbilical cord mesenchymal stem cells, and can solve the problems of uneven cell differentiation and difficulty in collection.

Inactive Publication Date: 2015-12-02
SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the defects of heterogeneous differentiation and difficult collection of cells produced by mass culture of mesenchymal stem cells in existing tissues, and to provide a mass culture method of umbilical cord mesenchymal stem cells

Method used

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Examples

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Effect test

Embodiment 1

[0032] S11, wash the umbilical cord with a length of 15 cm obtained from the operating room of the hospital twice with normal saline to ensure that the surface of the umbilical cord does not contain blood; remove both ends of the umbilical cord, and use normal saline to clean as much as possible the blood stains in the venous arteries of the umbilical cord;

[0033] S12 Use surgical scissors to cut the umbilical cord into small sections, each about 4 cm, remove the venous blood vessels and arterial blood vessels in turn, separate Wharton's jelly, put it in a petri dish and wash it with normal saline;

[0034] S13, cut Wharton's jelly into about 2-3mm 3 The small pieces of finely divided tissue should be kept as uniform as possible.

[0035] S21, inoculate the shredded Wharton's jelly into a T-75 culture flask, and make 5 repeated controls;

[0036] S22, after adding the complete medium to each culture bottle in step S21, put the culture bottle horizontally into the incubator ...

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Abstract

The invention provides a mass culture method of umbilical cord mesenchymal stem cells. The method comprises the following steps: obtaining umbilical cord Wharton's jelly; carrying out P0 generation culture on the umbilical cord Wharton's jelly until the cell fusion degree is 40-50%, and carrying out P1 subculture until the cell fusion degree is 85-95%; carrying out inoculating culture on P1 generation cells in a plate medium, and separating suspension cells non-adhered to the plate medium; and carrying out P2 subculture on the suspension cells in a medium containing a leukemia inhibition factor and a fibroblast-like growth factor. The above tissue adherent culture method allows adherence screening to be carried out through using a tissue culture plate after direct culture of P0 cells with low polytropy in early stage into P1 generation in order to maintain the uniformity of the cell differentiation degree; and the leukemia inhibition factor and the fibroblast-like growth factor are used to increase the adherence performance of the cells in the P2 generation culture process and inhibit differentiation, so the obtained umbilical cord mesenchymal stem cells have the advantages of low differentiation degree, trending to uniformity of the differentiation degree, strong adherence capability and collection facilitation.

Description

technical field [0001] The invention belongs to the field of stem cell preparation, and in particular relates to a mass culture method of umbilical cord mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells, MSCs) have the potential of multilineage differentiation under certain conditions, and are an important source of seed cells in tissue engineering research. Because umbilical cord Wharton's jelly (or also called Wharton's jelly, Walton's jelly) contains abundant mesenchymal stem cells, the mass preparation of mesenchymal stem cells is mostly obtained by separating and culturing from umbilical cord, and the preparation method is generally For the tissue block adherence method and enzyme digestion method. The usual enzymatic digestion method uses collagenase and trypsin to digest the umbilical cord Wharton's jelly tissue and then culture it in large quantities. In this method, trypsin affects the cytoskeleton by dissociating ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 曾宪卓黄海军
Owner SHEN ZHEN ISTEM REGENERATIVE MEDICINE SCI TECH CO LTD
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