Method for efficientlyobtaining mesenchymal stem cells from umbilical cord tissues

Abstract

The invention relates to amethod for efficientlyobtaining mesenchymal stem cells from umbilical cord tissues and aims at solving the problems of the long obtaining time and low efficiency of an existing method. The method comprises the steps that primary isolation of umbilical cord mesenchymal stem cells is performed: cleaned and cut umbilical cord tissue blocks are predigested by using trypsin and then digestive juice is removed, repeated flushing is performed with DPBS, then digestion is performed on a water bath shaker of 37 DEG C by using collagenase I digestive juice, a complete culture solution is added to the filtrate filtered by a cell strainer to terminate digestive centrifugal separation so as to obtain primary mesenchymal stem cell; residual tissue blocks are collected after filtration, and re-digestion is performed to obtain primary mesenchymal stem cells; the mesenchymal stem cells are inoculated in the complete culture solution, and standing and culture is performed in 5% CO2 culture box at the temperature of 37 DEG C; a culture solution is replaced for one time at intervals of several days; culture multiplication is performed: when the primary cells grow to be 80% and converge, TrypLE heavy suspension cells are subjected to culture multiplication in a serum-free culture solution. The method has the advantages of efficiently obtaining mesenchymal stem cells, and the obtained mesenchymal stem cells have highcell activity and differentiation potential.

Description

technical field [0001] The invention relates to a method for obtaining umbilical cord mesenchymal stem cells, in particular to a method for efficiently obtaining mesenchymal stem cells from umbilical cord tissue. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of stem cells, derived from the mesoderm in the early stage of development, have high differentiation potential, and are widely found in connective tissues such as bone marrow, fat, and umbilical cord blood in the human body and in the mesenchyme of organs. It has the potential of self-renewal, high proliferation and multi-directional differentiation. Under specific induction conditions in vivo and in vitro, it can differentiate into fat, bone, cartilage, muscle, cardiac muscle, endothelial and other tissue cells. With differentiation potential, it has gradually become a cell source of great practical value in the field of tissue engineering and gene therapy, and has attracted more and more atten...

AI Technical Summary

Problems solved by technology

The tissue block adhesion method mainly uses the adhesion and adhesion ability of MSCs to separate and purify MSCs cells during the adherent culture process, but the tissue block adhesion method needs to cut the umbilical cord tissue, and the mechanical external force will damage the tissue cells; The umbilical cord is a tissue rich in collagen and glucosamine, so the umbilical cord tissue is usually digested with collagenase to obtain MSCs. Enzymatic digestion can obtain a more complete cell structure and more MSCs, but most enzyme digestion methods take more time , the enzyme concentration and digestion time are very important for the isolation and culture of MSCs

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