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Method for efficiently separating umbilical cord mesenchymal stem cells

A technology of stem cells and umbilical cords, applied in the field of biological sciences, can solve the problems of isolated cell contamination, bacteria and other microorganisms that cannot be completely removed, and achieve the effects of small cell damage, good use effect, and guaranteed purity and vitality

Inactive Publication Date: 2015-08-26
GUIZHOU BEIKE FACTORR BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the umbilical cord is cleaned before the process of isolating umbilical cord mesenchymal stem cells, this cannot completely remove bacteria and other microorganisms carried on the surface of the umbilical cord, and the process of mechanically breaking the umbilical cord will cause the isolated cells to be polluted

Method used

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  • Method for efficiently separating umbilical cord mesenchymal stem cells
  • Method for efficiently separating umbilical cord mesenchymal stem cells

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Embodiment Construction

[0017] Embodiments of the present invention: a method for efficiently isolating umbilical cord mesenchymal stem cells, comprising the following steps:

[0018] 1) Separation of Wharton's jelly: Pour out the collection solution in the collection bottle, and only keep the umbilical cord in the collection bottle; add 75% alcohol by mass to completely submerge the umbilical cord and soak for 2 minutes; then transfer the umbilical cord to the culture medium. Wash twice with normal saline until the surface of the umbilical cord is free of blood; remove both ends of the cleaned umbilical cord, remove arteries, veins and amniotic membrane, separate Wharton's jelly, put it in a petri dish and use physiological Wash with salt water; cut Wharton's jelly into 1-4mm after cleaning 3 and transferred to a 50ml centrifuge tube;

[0019] 2) Inoculate the shredded Wharton's jelly into 2 T-75 culture flasks, no less than 1.0g in each flask, then add 9ml of complete medium to each culture flask,...

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Abstract

The invention provides a method for efficiently separating umbilical cord mesenchymal stem cells. Wharton jelly is directly separated from umbilical cord, so that parenchyma cell contamination is avoided, multi-time passage is not required, and the cell purity and activity are ensured; contamination in an umbilical cord separation process can be effectively alleviated, and the separation cost can be reduced. The umbilical cord does not need to be mechanically crushed in the separation process, so that the damage of mechanical shear force to the cells is reduced and the viability of the obtained cells is higher; heterologous digestive enzymes and heterologous serum are not adopted in the separation process, so that the obtained cells do not contain heterologous proteins and are also safer, the needs of clinical use can be met, and a technical guarantee is provided for establishing an umbilical cord mesenchymal stem cell library; a tissue block method is adopted, is more convenient and quick and small in cell damage, and can protect the mesenchymal stem cells from being not damaged and enable the mesenchymal stem cells to have higher viability.

Description

technical field [0001] The invention relates to the technical field of biological sciences, in particular to a method for efficiently isolating umbilical cord mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (mesenchyma1 stem cells, MSC S ) is a kind of pluripotent stem cell derived from mesoderm and ectoderm in early development, and was originally found in bone marrow, because it has the following characteristics; 1 hematopoietic support. As the main cellular component of the hematopoietic microenvironment - stem / progenitor cells of the stromal cell lineage, MSC S Co-transplantation with hematopoietic stem cells can promote the engraftment of hematopoietic stem and progenitor cells. 2 immune regulation. MSC S It does not express CD80, CD86, HLA-DR and other co-stimulatory molecules, and can inhibit mixed lymphocyte reaction in vitro, which is of great significance for the prevention and treatment of graft-versus-host disease caused by alloge...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 边曙光
Owner GUIZHOU BEIKE FACTORR BIOTECH CO LTD
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