Preparation method for improving yield of umbilical cord derived mesenchymal stem cell primary cells

Abstract

The invention relates to the technical field of cell culture and cell separation and in particular discloses a preparation method for improving the yield of umbilical cord derived mesenchymal stem cell primary cells. On the basis of culturing and separating mesenchymal stem cells on a traditional Wharton's jelly tissue block in a disposable adherence manner, Wharton's jelly tissue blocks are finely collected at different time of a later period of culture and are repeatedly adhered; P0 generation mesenchymal stem cells can be continuously collected at different time. According to the method provided by the invention, the utilization rate of Wharton's jelly is maximized through a property that the mesenchymal stem cells continuously move out from Wharton's jelly tissues to grow; compared with Wharton's jelly tissue block disposable adherence culture, the obtained quantity of the P0 generation mesenchymal stem cells can be increased by two times at least; the culture density of the mesenchymal stem cells, the concentration of serum replacements in a culture medium and the culture time are subsequently controlled; when the cells are sub-cultured to a P3 generation, the obtained cell quantity can completely meet the requirements of cell storage and clinical transplantation.

Description

technical field [0001] The invention relates to the technical field of cell culture and cell separation, in particular to a preparation method for increasing the yield of primary cells of umbilical cord-derived mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (Mesenchymal Stem Cells, MSCs) are a special type of stem cells, derived from the mesoderm and ectoderm in the early stages of development, and belong to pluripotent stem cells in classification. Under specific induction conditions in vivo or in vitro, MSCs can differentiate into adipocytes, bone cells, chondrocytes, nerve cells, liver cells, cardiomyocytes, endothelial cells, etc. It can be used as an ideal seed cell for the repair of tissue and organ damage caused by aging and disease. Because of its multi-directional differentiation potential, it can be widely used in clinical treatment of various diseases such as liver fibrosis and nerve injury. , cerebral palsy, myocardial infarction an...

AI Technical Summary

Problems solved by technology

The number of primary mesenchymal stem cells obtained, that is, the number of seed cells directly determines the number of high-generation cells obtained through subculture. Studies have shown that each centimeter of umbilical cord theoretically contains about 1.00×10 6 Mesenchymal stem cells, but in the actual preparation process of mesenchymal stem cells, the methods disclosed in the prior art (such as patent applications with publication numbers CN105420183A, CN105586309A, CN102154200B, and CN102660497B) all have more long shuttles around the tissue block. When developing mesenchymal stem cells, the tissue block is deducted and discarded, and only the cells that have dissociated and attached to the bottom of the culture container are expanded and passaged, but there are still many undissociated cells in the discarded tissue block. Mesenchymal stem cells

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