Method for rapidly obtaining and purifying human umbilical cord mesenchymal stem cells

Abstract

The invention relates to a method for rapidly obtaining and purifying human umbilical cord mesenchymal stem cells. The method comprises the following steps: (1) under a sterile condition, collecting the umbilical tissues of human newborns, washing the umbilical tissues by a sterile PBS liquid to remove blood and stains, and removing blood vessels from umbilical tissues by a mechanical separation method to obtain Wharton's jelly blocks; (2) cutting the Wharton's jelly blocks obtained in the step one into small blocks by a sterile scissor, re-suspending the small blocks in a cell culture fluid, inoculating the small blocks in a culture dish, and incubating the culture dish in an incubator to obtain adherent cells; and (3) discarding the culture liquid and suspended cells in the culture dish in the step (2), using composite collagenase to carry out differential digestion to digest adherent cells, and carrying out PBS washing and re-suspending to obtain cell suspension: P1 generation hUMSCs. The method has the advantages that during the whole process, all operation is performed by using antibiotic-free liquids, the toxin of antibiotic to cells is avoided, and thus the proliferation activity of hUMSCs is not influenced.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a method for obtaining and purifying stem cells. Specifically, it relates to a method for rapidly obtaining human umbilical cord mesenchymal stem cells and obtaining highly purified stem cells in a short period of time. Background technique [0002] Mesenchymal stem cells (marrow stromal cells, MSCs) are widely present in the interstitium of tissues and organs of the body, and have the ability of self-proliferation and multi-lineage differentiation. A large number of literatures have reported that MSCs can differentiate into terminal cells belonging to mesoderm, endoderm and even ectoderm under different conditions. Therefore, it is widely used in cell biology, tissue engineering and other research. [0003] In tissue engineering research, the source of MSCs has been the bottleneck of its development. Among the MSCs used in the past, the number of MSCs derived from adult cells is lim...

AI Technical Summary

Problems solved by technology

Among them, more stem cells can be obtained by enzymatic digestion of gel blocks, but this method requires the gel blocks to be soaked in digestive fluid for a long time, which has a great impact on the vitality of stem cells, and the ability of cell adhesion and proliferation often fails to meet the requirements; Although the adherence method to obtain hUMSCs has low requirements and is easy to promote, the passage cycle required to improve the purity of stem cells is long, and the speed often cannot meet the requirements.

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