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Preparation method of placenta decidua mesenchymal stem cells

A technology of stem cells and placenta, applied in the field of biomedicine, can solve the problems of low production efficiency and difficulty in large-scale production, and achieve the effects of large-scale production, thorough digestion and favorable growth

Pending Publication Date: 2020-12-01
江苏育瑞康生物科技有限公司
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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for preparing placental decidua mesenchymal stem cells and its preparation method, to solve the existing low production efficiency of mesenchymal stem cells isolated from placenta by tissue block adhesion method and enzyme digestion method , resulting in the difficulty of achieving mass production

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  • Preparation method of placenta decidua mesenchymal stem cells

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Embodiment 1

[0020] like figure 1 As shown, the preparation method of placental decidual mesenchymal stem cells comprises the following steps:

[0021] S1. Take out the placental decidua material containing mesenchymal stem cells from the sampling bottle, put it into a sterile tray containing 75% alcohol, rinse and soak for 1 min.

[0022] S2. Washing the placental decidua material with physiological saline to remove alcohol, and then removing the epithelial layer and calcified layer on both sides.

[0023] S3. Washing with D-Hanks solution, removing the blood vessels on each small block and extruding the blood in the blood vessels.

[0024] S4. Use sterile scissors to cut the placental decidua material into small pieces with a length of 1-2 cm, wash with normal saline until there is no blood color, and then cut the small pieces of the placental decidua material into small pieces with a length of 1-3 mm.

[0025] S5. Digest the obtained tissue block with a combined digestive enzyme, and ...

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Abstract

The invention provides a preparation method of placenta decidua mesenchymal stem cells. The preparation method comprises the following steps: s1, putting a placenta decidua material into a sterile tray containing 75% alcohol; s2, cleaning the placenta decidua material with normal saline to remove alcohol; s3, conducting cleaning through a D-Hanks solution, removing blood vessels on each small block and squeezing out blood in the blood vessels; s4, cutting the placenta decidua material into small pieces by using scissors, cleaning the small pieces by using normal saline, and then cutting the small pieces into tissue blocks; s5, performing digesting by using a combined digestive enzyme; s6, diluting the tissue mixture by using PBS (Phosphate Buffer Solution) with the same volume, and takingsupernate suspension; s7, centrifuging the supernate suspension to obtain a precipitate, resuspending the precipitate with an erythrocyte lysis solution, performing standing for lysis, performing centrifuging to obtain a precipitate, resuspending the precipitate with a culture solution, and performing inoculating to a culture dish; and s8, carrying out serum-free amplification passage on the precipitate in the culture dish to obtain passage clinical-grade placenta mesenchymal stem cells. The whole culture period is short, the efficiency is high and the stability is good.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a preparation method of placental decidua mesenchymal stem cells. Background technique [0002] Mesenchymal Stem Cells (Mesenchymal Stem Cells, MSCs) are mainly derived from the mesoderm in early embryonic development, which have the characteristics of self-renewal, multidirectional differentiation and immune regulation. Under certain conditions, it can differentiate into multiple functional cells like APSC pluripotent cells. The main function of placental mesenchymal stem cells is to have an immunoregulatory effect. Through the negative immune regulatory function, the hyperactive immune response of the body can be suppressed, and the immune function of the body can be restored to balance, so that it can be used to treat immune rejection after hematopoietic stem cell transplantation and cloning. Disease, lupus erythematosus, scleroderma and other autoimmune disea...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2509/00
Inventor 王建波
Owner 江苏育瑞康生物科技有限公司
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