Method for separation and purification of human embryonic trophoblast cells and placental mesenchymal stem cells

Abstract

The invention discloses a method for separating and purifying human embryonic trophoblast cells and placental mesenchymal stem cells. The method comprises the following steps: (1) placenta treatment;(2) pre-separation of a cell suspension; (3) flow adsorption separation; (4) desorption and separation of the placental mesenchymal stem cells; and (5) purification of the human embryonic trophoblastcells. In the invention, an antibody protein is planted on the outer surface of a temperature-sensitive chitosan copolymer membrane, the surface molecules of the placental mesenchymal stem cells are recognized by the antibody to achieve the purpose of specific binding, and a decreasing flow rate circulating manner is adopted to reserve the placental mesenchymal stem cells on the specific temperature-sensitive chitosan copolymer membrane. The desorption and separation of the placental mesenchymal stem cells and a protein antibody carried out by cooling and eluent addition. The method has the advantages of mild separation way, high cell survival rate and high purity.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for separating and purifying human embryo trophoblast cells and placental mesenchymal stem cells. Background technique [0002] 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. [0003] The conventional methods currently used for trophoblast cells and placental mesenchymal stem cells include the following four: [0004] (1) Adhesive separation method. This method mainly utilizes the characteristics of stem cells to adhere to the wall in plastic culture flasks to separate stem cells from other...

AI Technical Summary

Problems solved by technology

[0008] The thermosensitive chitosan copolymer film has good temperature-sensitive characteristics, so that it can achieve the purpose of artificially controlled cell adsorption and desorption only by changing the temperature without the need for enzymatic hydrolysis. Compared with immunomagnetic The bead method is gentler, less damage to the cells, and the success rate of the cells is high, and it is easier to obtain a large number of primary cells at one time. There are few reports on the separation and purification of cells by combining them with specific antibody proteins.