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Preparation method for mesenchymal stem cells from placental decidua basalis

A technology of placental decidual basal mesenchymal stem cells, applied in the field of placental decidua basal mesenchymal stem cells, can solve the problems of prolonging cell culture time, cell waste, increasing the difficulty of cell expansion, etc., and shorten the time of cell culture Effects of time and culture cycle, increasing utilization rate, avoiding cell quality decline and functional morphological changes

Inactive Publication Date: 2015-06-10
希瑞干细胞科技有限公司
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the enzymatic digestion method is commonly used to obtain mesenchymal stem cells from placenta and umbilical cord. Usually, collagenase is used for 2 to 3 hours to digest and then trypsin is added. This enzymatic digestion method will result in a small amount of mesenchymal stem cells due to incomplete digestion. Unseparated cells are wasted, increasing the difficulty of cell expansion and prolonging cell culture time

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  • Preparation method for mesenchymal stem cells from placental decidua basalis
  • Preparation method for mesenchymal stem cells from placental decidua basalis
  • Preparation method for mesenchymal stem cells from placental decidua basalis

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[0029] The beneficial effect of the present invention will be further explained by comparing the preparation methods of several decidua mesenchymal stem cells at the base of the placenta. Several methods for preparing placental decidua mesenchymal stem cells include tissue block attachment method, trypsin digestion method, trypsin plus collagenase direct digestion method and the preparation method of the present invention. Among them, type Ⅰ collagenase, trypsin, and DMEM-F12 medium were all purchased from GIBCO Company. The comparison operation includes the following steps:

[0030] S101. The decidua basalis is stripped from the fresh term placenta tissue, and the decidua basalis tissue is washed with normal saline.

[0031] S102. Drain the normal saline on the surface of the decidua basalis tissue as much as possible, and cut it into pieces in a glass dish, and the volume of the tissue pieces is 1 mm. 3 Left and right, and then equally divided into A, B, C, D4 groups.

[...

Embodiment 1

[0063] The preparation method of the placental base decidua mesenchymal stem cells of the present embodiment comprises the following steps:

[0064] S201. The decidua basalis is stripped from the fresh full-term placental tissue, and the decidua basalis tissue is washed with normal saline. Drain the normal saline on the surface of the decidua basalis tissue as much as possible, cut it into pieces in a glass dish, and the volume of the tissue pieces is 1mm 3 about.

[0065] S202. Add type I collagenase to the decidua basal tissue block and digest at 37° C. for 18 hours. The amount of type Ⅰ collagenase added was twice the volume of the decidua basal tissue block.

[0066] S203, add trypsin and continue to digest at 37° C. for 20 minutes, then filter and centrifuge to obtain placental decidua mesenchymal stem cells. The amount of trypsin added was 0.2 times the total volume of decidual tissue pieces digested with type Ⅰ collagenase.

[0067] S204, the decidua basalis tissue ...

Embodiment 2

[0071] The preparation method of the placental base decidua mesenchymal stem cells of the present embodiment comprises the following steps:

[0072] S201. The decidua basalis is stripped from the fresh full-term placental tissue, and the decidua basalis tissue is washed with normal saline. Drain the normal saline on the surface of the decidua basalis tissue as much as possible, cut it into pieces in a glass dish, and the volume of the tissue pieces is 1mm 3 about.

[0073]S202. Add type I collagenase to the decidual tissue block and digest at 37° C. for 18.5 hours. The amount of type Ⅰ collagenase added was twice the volume of the decidua basal tissue block.

[0074] S203, add trypsin and continue to digest at 37° C. for 30 minutes, then filter and centrifuge to obtain placental decidua mesenchymal stem cells. The amount of trypsin added was 0.25 times the total volume of decidual tissue pieces digested with type Ⅰ collagenase.

[0075] S204, the decidua basalis tissue blo...

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Abstract

The invention discloses a preparation method for mesenchymal stem cells from placental decidua basalis. The preparation method includes the following steps that after I-type collagenases are added to decidua basalis tissue blocks to digest for 18 h-20 h, pancreatic enzymes are added to continue to digest, filtration and centrifugation are then conducted, and the mesenchymal stem cells from placental decidua basalis are obtained; the obtained mesenchymal stem cells from placental decidua basalis are cultured and amplified. The I-type collagenases digest the decidua basalis tissue blocks for a long time so that a great number of mesenchymal stem cells can be obtained, the utilization rate of the decidua basalis tissue blocks is increased, cell culture time and the cell culture period are shortened, cell quality reduction and functional form change caused by repeated cell replication are avoided, and a good guarantee is provided for the clinical mesenchymal stem cells from placental decidua basalis.

Description

technical field [0001] The invention relates to the preparation of mesenchymal stem cells, in particular to a method for preparing placental decidua mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells derived from mesoderm and ectoderm in early development. They were originally found in bone marrow because of their following characteristics: 1) Hematopoietic support. As the main cell component of the hematopoietic microenvironment—the stem / progenitor cells of the stromal cell line, co-transplantation of MSCs and hematopoietic stem cells can promote the implantation of hematopoietic stem and progenitor cells; 2) immune regulation. MSCs do not express CD34, CD45, HLA-DR and other co-stimulatory molecules, and can inhibit mixed lymphocyte reaction in vitro, which is of great significance for the prevention and treatment of graft-versus-host disease caused by allogeneic hematopoietic cell transplantation and th...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 姚静张瑞婷杨莉莉
Owner 希瑞干细胞科技有限公司
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