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Method for inducing bone marrow mesenchymal stem cells to be differentiated into neuron-like cells

A technique for differentiation of neuron-like cells and stem cells, which is applied in the biological field and can solve problems such as the lack of physiological characteristics of nerve cells

Inactive Publication Date: 2013-04-03
陆华
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Problems solved by technology

However, these studies mostly focus on the morphological characteristics of neurons and the expression of neuron markers after induction, but lack evidence of physiological characteristics of neurons. Therefore, many scholars believe that these cells are called "neuron-like cells". more appropriate

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  • Method for inducing bone marrow mesenchymal stem cells to be differentiated into neuron-like cells

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[0014] like figure 1 Shown is a schematic flow chart of the steps of the all-trans retinoic acid combined with cytokine induction method of bone marrow mesenchymal stem cells differentiated into neuron-like cells of the present invention, the method of the embodiment includes the following steps:

[0015] S1. Isolation and culture of cells: All operations were performed under sterile conditions, and the bone marrow was collected from cancellous bone fragments during hip joint surgery or open surgery on lower limb bones. The bone marrow suspension collected by anticoagulation with heparin was diluted with D-Hanks solution (V:V, 1:1), slowly added to half the volume of Ficoll separation medium (lymphocyte separation medium), centrifuged at 600g for 15min, and the interfacial layer cells were collected. D-Hanks solution was washed once, and the cell concentration was adjusted to 1×10 5 / ml, inoculated at 25cm 2 The medium is DMEM-LG (low sugar DMEM medium) + 1% Penicillin-Strep...

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Abstract

The invention provides a method for inducing bone marrow mesenchymal stem cells to be differentiated into neuron-like cells. The method comprises the steps of cultured cell separation, mesenchymal stem cell morphology observation, flow cytometry antigen presentation detection, differentiation experiment induction and cell immunofluorescence staining. According to the method, the bone marrow mesenchymal stem cells are induced to be differentiated into the neuron-like cells by using all-trans retinoic acids, basic fibroblast growth factors and epidermal growth factors, so that the neuron-like cells not only have the typical morphology of nerve cells, but also express the neuron-specific enolase of neuron marker antigens and the glial fibrillary acidic protein of astrocyte marker antigens; and the bone marrow mesenchymal stem cells maintain the capability that the bone marrow mesenchymal stem cells can be differentiated into non-mesenchymal cells in a germ-layer-crossing manner, and can be differentiated into the neuron-like cells in the germ-layer-crossing manner, so that the neuron-like cells are expected to become seed cells for nerve cell replacement therapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for induction of bone marrow mesenchymal stem cells into neuron-like cells combined with all-trans retinoic acid combined with cytokines. Background technique [0002] Although bone marrow mesenchymal stem cells (BMSCs) transplantation has made some progress in the treatment of ischemic brain injury, the conversion rate of nerve cells after BMSCs transplantation can be as high as 80% in vitro, but only 3% to 10% in vivo. In the in vivo environment, the rate of differentiation into glial cells is large and the rate of differentiation into neuron-like cells is small, which will undoubtedly bring unfavorable factors to the recovery of nervous system function. If bone marrow stromal cells can be differentiated into neuron-like cells in vitro, cell transplantation will have a multiplier effect. [0003] Studies on the differentiation of mesoderm-derived BMSCs to ectoderm-derived...

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Application Information

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IPC IPC(8): C12N5/079
Inventor 陆华
Owner 陆华
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