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Inducing method for differentiating umbilical cord mesenchymal stem cells into neural stem cells

A technology of neural stem cells and mesenchymal stem cells, applied in animal cells, nervous system cells, vertebrate cells, etc., can solve problems such as ethics, safe cell sources and limited quantity

Inactive Publication Date: 2013-04-10
陆华
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Problems solved by technology

However, ethics, safety issues, and the limited source and quantity of cells limit the application of neural stem cell transplantation to a certain extent.

Method used

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  • Inducing method for differentiating umbilical cord mesenchymal stem cells into neural stem cells

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Embodiment Construction

[0013] Such as figure 1 Shown is a schematic flow chart of the steps of the method for inducing differentiation of umbilical cord mesenchymal stem cells into neural stem cells of the present invention, the method of the embodiment includes the following steps:

[0014] S1. Isolation and primary culture of UC-MSCs:

[0015] Under sterile conditions, the 10cm umbilical cord of a healthy fetus with full-term pregnancy by cesarean section was fully washed with D-Hanks solution containing 1% double antibody and 250ug / mL amphotericin, and the two umbilical cords were inserted into two umbilical cords with 20mL syringes respectively. Repeated washing in the vein and an umbilical artery to remove the residual blood in the umbilical vein and artery; tear off the amniotic membrane on the surface of the umbilical cord to obtain mesenchymal tissue, and cut the umbilical cord tissue into 1mm pieces with tissue scissors 3 Large or small tissue pieces, move the tissue pieces (about 30-40ml)...

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Abstract

The invention provides an inducing method for differentiating umbilical cord mesenchymal stem cells (UC-MSCs) into neural stem cells. The inducing method comprises the following steps: carrying out separation and primary culture of UC-MSCs, subculture and amplification of UC-MSCs, induction in vitro of UC-MSC to be transformed into neural stem cells, and differentiation culture of neural stem cells and detection by an immumofluorescence method. The inducing method provided by the invention uses all-transretinoic acid combined with alkaline fibroblast growth factors (bFGF) and epidermal growth factor (EGF) to induce the UC-MSCs to be transformed into neural stem cells, wherein the UC-MSCs still have stronger activity after repeated passage by division growth. The transretinoic acid is combined with cytokines to induce the UC-MSCs to be converted into the neural stem cells, so that the mesenchymal cells are differentiated into non mesenchymal cells cross germinal layer, and the mesenchymal cells probability become more ideal seed cells in the future clinical application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing differentiation of umbilical cord mesenchymal stem cells into neural stem cells by using all-trans retinoic acid combined with cytokines. Background technique [0002] At present, neural stem cells are mainly induced from embryonic stem cells or directly isolated and cultured from the central nervous system of developing and adult mammals. However, ethics, safety issues, and the limited source and quantity of cells limit the application of neural stem cell transplantation to a certain extent. Therefore, it is necessary to find other ways to obtain neural stem cells to overcome these limitations. The research on the transformation of mesoderm-derived mesenchymal stem cells into ectoderm-derived neural stem cells is in its infancy. Umbilical cord mesenchymal stem cells (UC-MSCs) come from newborns, which are convenient to obtain, easy to expand in vitro, safe and...

Claims

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Application Information

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IPC IPC(8): C12N5/0797C12N5/0775
Inventor 陆华
Owner 陆华
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