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

A technology for nourishing cells and stem cells, applied in the field of biomedicine, can solve the problems of difficulty in obtaining large quantities and purity, complex placental cell components, etc., and achieve the effects of ideal cell purity and vitality, low cost, and small cell damage.

Inactive Publication Date: 2015-11-25
大连金玛健康产业发展有限公司
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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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  • Method for separating and purifying human embryo trophoblast and placental mesenchymal stem cells
  • Method for separating and purifying human embryo trophoblast and placental mesenchymal stem cells
  • Method for separating and purifying human embryo trophoblast and placental mesenchymal stem cells

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Embodiment Construction

[0023] The technical scheme of the present invention will be described in further detail below in conjunction with specific tests carried out in the laboratory, but the present invention is not limited to the specific examples listed below.

[0024] A method for isolating and purifying human embryonic trophoblast cells and placental mesenchymal stem cells, comprising the following steps:

[0025] (1) Treatment of placenta specimens: Under aseptic conditions, remove the amnion from the delivered placenta, cut off the 2.5mm tissue on the maternal surface of the placenta, cut off the placenta leaflets, weigh 50g, and soak them in the preservation solution containing antibacterial peptides. The peptide preservation solution is prepared by adding 55 micrograms / ml polylysine to 55 micrograms / ml gallate EGCG. After soaking for a predetermined time, cut the cut placental leaflets with surgical scissors and wash with normal saline. Wash the blood stains repeatedly until the washing liq...

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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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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073C12N5/0775
Inventor 何静
Owner 大连金玛健康产业发展有限公司
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