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Method for inducing differentiation from human umbilical cord mesenchymai stem cells (hucMSCs) into neural cells

A technology of stem cells and nerve cells, applied in the field of cell biology

Inactive Publication Date: 2011-09-21
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Abstract
  • Description
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Problems solved by technology

So far, there is no report on the induction of HUCMSCs into neuron-like cells by chemical or co-culture methods and their application in the research of nerve injury repair

Method used

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  • Method for inducing differentiation from human umbilical cord mesenchymai stem cells (hucMSCs) into neural cells
  • Method for inducing differentiation from human umbilical cord mesenchymai stem cells (hucMSCs) into neural cells
  • Method for inducing differentiation from human umbilical cord mesenchymai stem cells (hucMSCs) into neural cells

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Embodiment Construction

[0026] Experimental steps:

[0027] 1. Materials and methods

[0028] The umbilical cords were obtained from full-term caesarean section infants in the Department of Obstetrics and Gynecology, Shanghai First People's Hospital, with the authorization and consent of their parents.

[0029] 2. Main reagents and factors

[0030] Ten 2-week-old C57BL / 6 mice (Shanghai Experimental Animal Breeding Center, Chinese Academy of Sciences), DMEM medium (Gibco, USA), complex collagenase NB4 (Serva, Germany), fetal bovine serum (FBS: Hyclone, Australia), phosphoric acid Salt buffer (PBS, PH7.2), Schwann cell culture medium (SCCM, Schwann cells culture medium containing 10ngheregulin-β-1, 2uM forsklin, 10% FBS, 100u / ml penicillin, 100u / ml streptomycin and high sugar DMEM). Rabbit anti-human NF-200, B-tubulin III, GFAP, Nestin (Milipore, USA) flow cytometry antibodies were purchased from (BD, USA).

[0031] 3. Isolation of umbilical cord mesenchymal cells

[0032] Remove the umbilical cor...

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Abstract

The invention belongs to the field of cell biology and relates to a method for inducing differentiation from human umbilical cord mesenchymai stem cells (hucMSCs) into neural cells. The method comprises that HUCMSCs and Schwann cells are putted in Transwell with apertures of 0.4 microns to be co-cultured separately; a Schwann cell culture medium is utilized as a cell culture medium and half or all of the cell culture medium is replaced every three days; and then neural cells can be obtained after two weeks of culturing. Through a separate co-culturing method utilized by the invention, an umbilical cord can be digested by a mixture of composite collagenase NB4 and hyaluronidase and thus a large amount of HUCMSCs can be separated. High purity HUCMSCs can be obtained in third generation passage cells. In the invention, HUCMSCs are induced and differentiate into neural cells, wherein a cell differentiation rate is over 70%. Expressing rate ranges of specific markers, such as NF-200, nestin, Betar-III-tubulin, etc., of neural cells are from 70 to 80%.

Description

technical field [0001] The invention belongs to the field of cell biology and relates to a method for stably inducing human umbilical cord mesenchymal stem cells. Background technique [0002] The repair of nervous system trauma is still a big problem to be solved in clinical practice. Autologous nerve transplantation, which is the gold standard for repairing nerve defects, has the disadvantages of limited sources and left sensory impairment at the donor site. [0003] Stem cells have the characteristics of abundant sources, easy isolation and culture, strong in vitro proliferation ability, and multi-directional differentiation potential. In recent years, they have become a hot spot in the research of seed cells for tissue engineering products. Studies have found that embryonic stem cells (ESCs), neural stem cells (NSCs), bone marrow stromal stem cells (BMSCs), and umbilical cord blood stem cells all have the potential to differentiate into nerve cells, and can improve thei...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 沈尊理祝加学秦金保
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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