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Method for inducing neural differentiation of stem cells

A technology of stem cells and nerves, applied in the field of cell engineering, can solve the problems of short survival time and prone to apoptosis of nerve-like cells, and achieve high differentiation rate and survival rate

Inactive Publication Date: 2012-07-04
YEACELL BIOTECH INC
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Problems solved by technology

Among them, the Woodbury chemical induction method is currently a commonly used method for inducing the neural differentiation of rat MSCs. Under the culture conditions of various chemical inducers and growth factors in vitro, the conversion rate of MSCs to neural-like cells is as high as 50%-90%, but the differentiation rate is as high as 50%-90%. Most of the post-induction cells only confirmed the morphology of nerve cells and the expression of surface markers, but there are few reports on whether they have mature nerve cell functions, and the induced nerve-like cells are prone to apoptosis and have a short survival time.

Method used

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  • Method for inducing neural differentiation of stem cells
  • Method for inducing neural differentiation of stem cells
  • Method for inducing neural differentiation of stem cells

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Embodiment 1

[0038] The first step is to isolate, culture and identify primary rat MSCs: take SPF grade 4-8 week old SD rats, take the femur under sterile conditions, absorb DMEM / F12 with a 5ml empty needle, repeatedly rinse the bone marrow cavity and dry dirt end, and centrifuge Remove the supernatant, add DMEM / F12 / 10%FBS / 1% penicillin and streptomycin complete medium by pipetting, press 3x10 6 The concentration of one / bottle was inoculated at 25cm 2 placed in a plastic culture bottle at 37°C, 5% CO 2 After culturing in a humidified incubator, discard the supernatant and unattached cells after 24 hours. After changing the medium, it can be seen that rMSCs grow in scattered cell colonies, and the individual cells are flat spindle-shaped, and the cells in the center of the colony are round. The cells at the edge of the colony are flattened and fusiform. After 4-5 days, the number of cells increased significantly, and the colonies gradually expanded. After 6 to 7 days, the cells basicall...

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Abstract

The invention relates to a method for inducing neural differentiation of stem cells. All-transretinoicacid (ATRA) serves as an inducer and a modified neuronal induction medium (MNM) serves as a culture medium used for treating the stem cells, so the neural differentiation of the stem cells can be promoted effectively, differentiation ratio and survival ratio are increased, neuron-like cells with the neuronal function are prepared, a good in-vitro model is provided for research on development of nerve cells, and powerful basis is provided for the stem cells applied to clinical treatment on nerve system diseases.

Description

technical field [0001] The invention relates to cell engineering technology, and relates to an in vitro induction method for promoting neural differentiation of stem cells. Background technique [0002] Refractory diseases of the nervous system are mostly caused by damage to the nervous system, which leads to the loss of a large number of neuron structures and functions. The traditional view is that nerve cells lack the ability to regenerate, cannot generate new neurons and establish synaptic connections, and nerve damage is irreversible. In recent years, studies have found that transplanting stem cells into damaged or injured brain tissue can generate new nerve cells and repair and improve the nervous system in terms of structure and function. Stem cells, especially adult stem cells, as a new type of biological treatment is a pioneering exploration in the history of medical development. [0003] Bone marrow mesenchymal stem cells (MSCs) are a type of stem cells that exist ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/0793
Inventor 李廷玉毕杨龚敏陈洁魏小平张贇
Owner YEACELL BIOTECH INC
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