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In-vitro culture and proliferation method for human mesenchymal stem cells

A bone marrow mesenchymal, in vitro culture technology, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of low stem cell growth purity, large cell damage, weak cell viability, etc., to achieve simple and easy operation. The effect of line, strong cell viability and simple method

Active Publication Date: 2013-12-04
奥思达干细胞有限公司
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AI Technical Summary

Problems solved by technology

The stem cells obtained by adherent culture method and Ficoll separation solution method grow faster but the purity is too low
The purity of the stem cells obtained by the Percoll separation method is better, but the growth rate is slower and some stem cells may be missing
③Flow cytometry screening method or immunomagnetic bead method: cells are separated according to different cell sizes or some special markers on the cell surface. weak cell viability
However, the serum-free medium is chemically limited, and it still changes during the culture process. Some substances may be lacking at the beginning of the culture, and then the product of the cells may accumulate, so that the composition of the medium changes. negative proliferative state

Method used

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  • In-vitro culture and proliferation method for human mesenchymal stem cells
  • In-vitro culture and proliferation method for human mesenchymal stem cells
  • In-vitro culture and proliferation method for human mesenchymal stem cells

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Embodiment Construction

[0033] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.

[0034] refer to Figure 4 As shown, the specific implementation of this embodiment is as follows:

[0035] 1) Preparation of bone marrow mesenchymal stem cell culture medium, the steps include:

[0036] a) Configure 950ml DMEM medium as the basic medium; b) Add 300mg L-glutamine, 50mg vitamin C, 15mU insulin, 10U erythropoietin and 10μg human stromal cell-derived factor-1 in turn; c) Add sodium hydroxide Adjust the pH to 7.0-7.2; d) Add 50ml of fetal bovine serum, filter and sterilize, and store at 4°C for later use.

[0037] 2) Cell culture

[0038] The primary cells of bone marrow mesenchymal stem cells were collected, digested with 0.25% trypsin to make a cell suspension, counted the number of cells, and adjusted the cell density to 2×105 cells / ml inoculated to the medium containing bone marrow mesenchymal stem cells; inoculate...

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Abstract

The invention discloses an in-vitro culture and proliferation method for human mesenchymal stem cells. A DMEM culture medium is taken as a basic culture medium and is sequentially added with L-glutamine, vitamin C, insulin and erythropoietin; then the PH value is adjusted by sodium hydroxide; finally, fetal calf serum is added and filtration sterilization is performed to obtain a new culture medium, and cells are cultured on the basis of the new culture medium. The method can remarkably improve proliferation quantity and cell activity of human mesenchymal stem cells, meanwhile keeps biological effects and multi-lineage differentiation, enhances the efficacy of repairing body histiocyte damage through cell transplantation, and can be promptly and effectively used in scientific researches and clinic cases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an in vitro culture and expansion method for human bone marrow mesenchymal stem cells, which can significantly increase the number and activity of cell expansion, and enhance the efficacy of cell transplantation to repair damage to human tissue cells. Background technique [0002] Human bone marrow mesenchymal stem cells, as a new kind of widely used seed cells, not only have the ability to proliferate, but also have the potential of multi-lineage differentiation. Under specific induction conditions, they can differentiate into bone cells or chondrocytes, neuron-like cells, Fat cells, muscle cells, nerve cells and other tissue cells. At the same time, it can secrete a variety of cytokines, growth factors and matrix molecules that promote the proliferation and maturation of hematopoietic stem cells; it is an important part of the bone marrow hematopoietic microenvironment, and can sup...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 周萱
Owner 奥思达干细胞有限公司
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