Method for separating and purifying human embryo trophoblast and placental mesenchymal stem cells

Abstract

The invention discloses a method for separating and purifying human embryo trophoblast and placental mesenchymal stem cells. The method includes: processing a placenta sample to obtain processed placenta tissues; digesting the tissues to obtain cell suspension; performing discontinuous Percoll density gradient separation to obtain trophoblast and placental mesenchymal stem cells; purifying the trophoblast; further separating and culturing the placental mesenchymal stem cells. The method has the advantages that HyQTase and DNAse I are used to jointly digest the tissues, and a large amount of trophoblast and placental mesenchymal stem cells with ideal cell purity and activity is obtained through the density gradient separation; digestion fragments, fibroblast and red blood cells are removed, the trophoblast and the placental mesenchymal stem cells are distinguished from each other, the purity of the trophoblast can reach 90%, the non-adherence upper layers of the placental mesenchymal stem cells are removed through a differential adhesion method, serum-free medium is added into a culture flask to continue the culture of the placental mesenchymal stem cells, and high purity of the placental mesenchymal stem cells is achieved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for separating and purifying human embryo trophoblast cells and placental mesenchymal stem cells. Background technique [0002] It is usually difficult to obtain a large number of trophoblast cells and placental mesenchymal stem cells with high purity by conventional separation methods; although the cells obtained by flow cytometry or magnetic bead sorting are of good purity, the cost is high. [0003] It is difficult to culture and passage trophoblasts in vitro, and many researchers adopt the scheme of direct treatment or intervention after separating cells, so obtaining a large number of trophoblasts at one time is very beneficial to the experiment. The abundance of placental mesenchymal stem cells in placental tissue is low, and a lot of placental tissue is needed to obtain a certain number of primary cells. [0004] In the prior art, the cell components of the p...

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Problems solved by technology

[0004] In the prior art, the cell components of the placenta are relatively complex, and the trophoblasts contained therein play an important role in the process of maternal-fetal immune tolerance. The placental mesenchymal stem cells have multi-directional differentiation potential and the characteristics of inhibiting lymphocyte proliferation. It is usually difficult to obtain the above two types of cells in large quantities and with high purity

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