Method for simultaneously extracting epithelial cells and mesenchymal stem cells from umbilical cord and placenta amnion tissues

Abstract

The invention relates to a method for simultaneously extracting epithelial cells and mesenchymal stem cells from umbilical cord and placenta amnion tissues. The method comprises the steps of (1) sampling umbilical cord and placenta amnia and inoculating; (2) performing amplifying and passage on mesenchymal stem cells and amnion epithelial cells in a culture flask; (3) identifying the mesenchymal stem cells and the amnion epithelial cells: 1) when culture cells is transferred to the third generation, identifying the purity of culture cells after trypsinization; 2) verifying the cell purity to be 90 percent or more, namely the standard is met, and a mixture of the epithelial cells and the mesenchymal stem cells is obtained. The method is low in cost, simple and rapid, umbilical cord and placenta amnia are cut into tiny pieces by using a tissue cutting machine, which is conducive to swimming out of the tissues after the cells are directly inoculated in the culture flask; the cost is greatly lowered due to the fact that no proteinase reagent is used, and the preparation time is reduced, so that large-scale cell preparation in short time is realized; the method uses cell culture mediums being added with growth factors and other nutrients, and serum is saved.

Description

technical field [0001] The invention belongs to the technical field of cell culture, in particular to a method for simultaneously extracting epithelial cells and mesenchymal stem cells from umbilical cord and placental amniotic tissue. Background technique [0002] The umbilical cord and placenta belong to the tissues surrounding the embryo, and the umbilical cord tissue is the source of mesenchymal stem cells (MSCs). The placenta is composed of placental amnion, chorion and decidua basalis. The placenta contains a large number of stem cells, and the placental amniotic epithelial cells (AECs) have the characteristics of various stem cells, some of which are subtotipotent stem cells, whose characteristics are very close to those of embryonic stem cells (ESCs). The placental mesenchymal stem cells exist under the amniotic epithelial cell layer. Amniotic membrane originates from the epithelial layer of the embryo, which occurs earlier than the inner, middle, and outer germ la...

AI Technical Summary

Problems solved by technology

[0008] Although the above method avoids the use of a large number of reagents including expensive collagenase, the tissue block to be planted is still relatively large, and the cells in the tissue cannot be moved out most effectively.

Moreover, manual cutting and laying is also very time-consuming. It is fashionable to use the amniotic membrane of the placenta, but it is not practical to use the umbilical cord because the amount of umbilical cord tissue is much larger and very hard.