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Method for inducing self-body stem cell differentiation to be nerve stem cell with liquor cerebrospinalis

A technology of bone marrow stem cells and neural stem cells, applied in the field of differentiation of bone marrow stem cells into neural stem cells, can solve the problems of long culture period and high cost, and achieve the effect of simple method, low cost and good effect

Inactive Publication Date: 2008-08-20
万美蓉 +1
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AI Technical Summary

Problems solved by technology

The disadvantages of this traditional method are: long cultivation period; expensive

Method used

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Experimental program
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Effect test

Embodiment

[0023] 1. Conventional low-sugar DMEM / F 12 Medium + 10% autologous serum, plus penicillin-streptomycin according to the usual amount;

[0024] 2. Take 15ml of bone marrow, separate the cell layer with lymphocyte separation medium, and wash three times with phosphate buffered saline PBS;

[0025] 3. Take 1×10 7 The cell suspension was inoculated in a 25ml polylysine-coated cell flask;

[0026] 4. Change half of the medium in three days, and change the full medium in five days;

[0027] 5. Two weeks later, the cells were expanded and passaged for 5 times to transform into bone marrow stem cells. (immunization histochemistry and CD34, CD44 positive rate 100%);

[0028] 6. Take 2×10 7 2ml of the cell suspension was injected into the subarachnoid space through a lumbar puncture, and 2-3ml of effusion was drawn before injection, and the serum was collected by centrifugation for equipment.

[0029] 7. The bone marrow stem cells passed down for 5 passages were covered with cereb...

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Abstract

The invention relates to a method of the induction and differentiation of the neural stem cells and the neural cells from bone marrow stromal cells which are cultured by autologous bone marrow, autologous serum and autologous cerebrospinal fluid. The method pertains to the methods for differentiating the bone marrow stem cells into the neural stem cells. A conventional low-sugar DMEM / F 12 culture medium is adopted, the autologous serum replaces the original fetal bovine serum, the autologous cerebrospinal fluid replaces the original additive and a stimulating factor, a small amount of autologous bone marrow is used for culturing more cells with the number that can meet the clinical needs. The method has the advantages of low cost and easy material selection, avoiding zoonosis and foreign protein exclusive reaction, and so on. The method of inducing the bone marrow stromal cells into the neural stem cells by using cerebrospinal fluid can effectively solve the shortcomings of the traditional methods.

Description

technical field [0001] The invention relates to a method for differentiating bone marrow stem cells into neural stem cells. Specifically, it is a method for culturing bone marrow mesenchymal cells from autologous bone marrow, autologous serum, and autologous cerebrospinal fluid to induce differentiation into neural stem cells and neuron cells. Background technique [0002] At present, bone marrow mesenchymal cells are cultured into bone marrow stem cells at home and abroad, usually with low-sugar DMEM plus 10% fetal bovine or with low-sugar DMEM / F 12 Serum-free culture with various nutrients added, the former harvests more cells but cannot avoid zoonotic diseases and rejection of foreign proteins; the latter harvests less cells and requires more bone marrow from the donor. The method of inducing bone marrow mesenchymal cells to become neural stem cells at home and abroad is: using neurobasal and B 27 As a culture medium, human epidermal stimulating factor (EGF) and basic f...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/0793C12N5/0797
Inventor 万美蓉曾因明
Owner 万美蓉
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