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Method for extracting original mesenchymal stem cells from placenta and serum-free amplification

A stem cell, serum-free technology, applied in the field of biomedicine, can solve problems such as adverse reactions and risks of patients, achieve the effect of original stem cell characteristics, improve product quality, and effectively expand cells

Inactive Publication Date: 2011-08-10
王泰华
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Problems solved by technology

[0003] The latest research has found that placental tissue has adherent cells similar to bone marrow mesenchymal stem cells; at the same time, the current system for in vitro culture of placental mesenchymal stem cells mainly uses a certain proportion (such as 10-20%) of fetal bovine Serum-based media (such as low-sugar DMEM, high-sugar DMEM, DMEM / F12, and low-serum medium MesenPRO RS) are used for adherent culture expansion, but these culture systems contain fetal bovine serum due to the addition of animal components. Complement components, which may cause adverse reactions in patients if transplanted and infused into patients, and have unforeseen dangers in the short term

Method used

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  • Method for extracting original mesenchymal stem cells from placenta and serum-free amplification
  • Method for extracting original mesenchymal stem cells from placenta and serum-free amplification
  • Method for extracting original mesenchymal stem cells from placenta and serum-free amplification

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Embodiment

[0017] (1) Obtaining human placenta-derived mesenchymal stem cells

[0018] Immediately place the fresh placenta in a sterile tray or container, use physiological buffer solution to remove the hyalinous mucus and maternal blood contamination on the surface of the placental tissue and umbilical cord, inject a balanced salt solution containing heparin anticoagulant and antibiotics through the umbilical artery, Bleed and start perfusion. After the initial perfusion, the placental leaflets were cut aseptically, and the placental tissue was digested with 0.1% type I collagenase (Gibco) in a 37°C water bath for 30 minutes while shaking slowly. Obtain the cell suspension, then pass the cell suspension through a cell sieve to remove tissue fragments, centrifuge and wash at 1500 rpm, resuspend the obtained cells in DMEM and centrifuge, inoculate the obtained cells into culture flasks, and incubate at 37°C for 5 %CO 2 , cultured under saturated humidity conditions. After 24 to 48 hou...

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Abstract

The invention belongs to the technical field of biomedicine and particularly discloses a method for extracting original mesenchymal stem cells from a placenta and serum-free amplification. The more original mesenchymal stem cells of the human placenta are extracted by using placenta tissues of the early aborted fetus, and a system of separating the mesenchymal stem cells from the placenta and amplification culture in vitro is built up. The genetic information and phenotype of the stem cells are more stable, the characteristics of the stem cells are more original, the cells can be effectively amplified, the product quality is improved, the stable and homogeneous mesenchymal stem cells of the placenta can be obtained, the method solves the problems that a large quantity of stem cells are needed for stem cell treatment, and the method can be used for treating various diseases.

Description

(1) Technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a method for extracting primitive mesenchymal stem cells from placenta and expanding them without serum. (2) Background technology [0002] Mesenchymal stem cells were originally discovered in the bone marrow, and have attracted increasing attention because of their multi-lineage differentiation potential, support for hematopoiesis and promotion of stem cell implantation, immune regulation, and self-replication. However, as far as the relatively mature bone marrow mesenchymal stem cells are concerned, their content is very low, accounting for only 1 / 10 of the monocytes in the bone marrow. 5 ~10 6 , and the collection of bone marrow brings great pain to patients, and with the increase of age, the number, proliferation and differentiation of bone marrow mesenchymal stem cells are significantly reduced, which limits its research and application, especially clinical applicat...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 王泰华李荣荣陈立才孙涛
Owner 王泰华
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