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Method for promoting proliferation of human umbilical cord mesenchymal stem cells and application thereof

A technology of mesenchymal stem cells and human umbilical cord, applied in the field of cell culture, can solve the problems of low yield and long expansion cycle, and achieve the effect of promoting proliferation, maintaining stem cell characteristics, cell vitality and strong proliferation ability

Pending Publication Date: 2020-04-17
上海林望生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, at present, umbilical cord mesenchymal stem cells have problems such as long expansion cycle and low yield.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] This embodiment discloses a method for promoting the proliferation of human umbilical cord mesenchymal stem cells, comprising the following steps:

[0036] S1: Human umbilical cord mesenchymal stem cells were isolated from human umbilical cord tissue;

[0037] S2: Rat colonic fibroblasts were isolated from rats;

[0038] S3: The human umbilical cord mesenchymal stem cells and the rat colonic fibroblasts were co-cultured in a transwell culture chamber and a cell culture plate, the upper chamber was inoculated with rat colonic fibroblasts, and the lower chamber was inoculated with human umbilical cord mesenchyme stem cell.

[0039] Specifically, the technical solution is as follows:

[0040] 1. Preparation of human umbilical cord mesenchymal stem cells

[0041] Get discarded newborn umbilical cords from the hospital, and cut human umbilical cord tissue pieces into 2-3mm within 24 hours 3 Small pieces, implanted into T75 culture bottles, density 20-25 pieces / bottle;

...

Embodiment 3

[0061] The human umbilical cord mesenchymal stem cells co-cultured in Example 1 were induced to differentiate into adipocytes, and it was found that after about one week of induction, the cell morphology changed from a long spindle to a short spindle, and showed cell aggregation. Small square-shaped cell aggregates. In about two weeks, lipid droplets with strong refraction can be seen in the cytoplasm, which were stained with Oil Red O, and the stained lipid droplets can be seen.

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PUM

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Abstract

The invention, which belongs to the field of cell culture, particularly relates to a method for promoting proliferation of human umbilical cord mesenchymal stem cells and application thereof. The method comprises the following steps: S1, separating human umbilical cord mesenchymal stem cells from human umbilical cord tissues; S2, performing in-vivo separation on a rat to obtain rat colonic fibroblasts; and S3, co-culturing the human umbilical cord mesenchymal stem cells and the rat colonic fibroblasts by using a transwell culture chamber and a cell culture plate, inoculating the rat colonic fibroblasts into an upper chamber, and inoculating the human umbilical cord mesenchymal stem cells into a lower chamber. The invention discloses a method for promoting proliferation of human umbilical cord mesenchymal stem cells by adopting a co-culture mode of rat colonic fibroblasts and the human umbilical cord mesenchymal stem cells for the first time. According to the method, more cells are obtained after proliferation, the cell viability and the proliferation capacity are high, the good proliferation capacity is still kept after the cells are cultured to P18 generations, and the form and good stem cell characteristics of the umbilical cord mesenchymal stem cells can be well kept.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to a method and application for promoting the proliferation of human umbilical cord mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (Mesenchymal stem cells, MSCs), also known as multipotential mesenchymal stromal cells, are non-hematopoietic adult stem cells that are widely distributed in tissues and have the ability of self-renewal and differentiation. MSCs were first isolated from bone marrow and subsequently obtained from other tissues and organs including hair follicles, tooth roots, periosteum, perichondrium, dermis, cord blood, umbilical cord, placenta, adipose muscle, lung, liver and spleen. Many studies have shown that MSCs cultured in vitro specifically express 113 transcripts and 17 proteins that hematopoietic cells do not express. According to the minimum standards for defining human MSCs proposed by the Mesenchymal and Tissue Stem Ce...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N2502/13C12N2501/11C12N2500/84C12N2509/00
Inventor 李志强张娜王钊林郝子娟燕小宁
Owner 上海林望生物科技有限公司