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Method for efficiently inducing umbilical cord mesenchymal stem cells to differentiate into Schwann-like cells

A technology of stem cells and Schwann cells, applied in the field of cell biology, can solve the problems of residual stem cells, patient trauma, application limitations, etc.

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

[0003] At present, it has been reported that embryonic stem cells, neural stem cells, bone marrow mesenchymal stem cells, and adipose stem cells are induced to differentiate into Schwann-like cells. and fat stem cells will bring new trauma to the patient, so its application is subject to certain limitations
Umbilical cord mesenchymal stem cells are stem cells isolated from fetal wastes and umbilical cords. They have strong self-proliferation ability, can be stably expanded for 80 generations without changing, and have greater proliferation ability than bone marrow mesenchymal stem cells. A variety of differentiation potentials, it has been proved that it can be induced to differentiate into bone, cartilage, fat, neurons and other cells, it is a stem cell between embryonic stem cells and adult stem cells, but the induction of differentiation into Schwann-like cells has not been reported

Method used

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  • Method for efficiently inducing umbilical cord mesenchymal stem cells to differentiate into Schwann-like cells

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Embodiment 1

[0023] Example 1 Method for Efficiently Inducing Human Umbilical Cord Mesenchymal Stem Cells to Differentiate into Schwann-like Cells

[0024] The operation steps are as follows:

[0025] (1) After culturing the umbilical cord mesenchymal stem cells to a subconfluent state, discard the old culture medium, and add a culture medium containing 1 mM β-mercaptoethanol to induce for 24 hours;

[0026] (2) Discard the induction solution described in step (1), wash three times with DMEM medium, add all-trans retinoic acid (RA) containing 35ng / ml for induction for 72 hours;

[0027] (3) Discard the induction solution described in step (2), wash three times with DMEM medium, add a composite induction solution [composed of 5ng / ml platelet-derived growth factor (PDGF), 10ng / ml basic fibroblast growth factor (bFGF ), 14μM forskolin (forskolin, FSK), 252ng / ml glial cell growth factor (GGF-2) or 200ng / ml nerve growth factor (heregulin-betal, HRG)] for 2 weeks, and change the solution every ...

Embodiment 2

[0030] Example 2 Method for Efficiently Inducing Human Umbilical Cord Mesenchymal Stem Cells to Differentiate into Schwann-like Cells

[0031] The operation steps are as follows:

[0032] (1) After culturing the umbilical cord mesenchymal stem cells to a subconfluent state, discard the old culture medium, and add a culture medium containing 1 mM β-mercaptoethanol for induction for 20 hours;

[0033] (2) Discard the induction solution described in step (1), wash it three times with DMEM medium, and add 35ng / ml all-trans retinoic acid (RA) for induction for 60 hours;

[0034] (3) Discard the induction solution described in step (2), wash 2 times with DMEM medium, add composite induction solution [by 6ng / ml platelet-derived growth factor (PDGF), 9 / ml basic fibroblast growth factor ( bFGF), 13 μM forskolin (forskolin, FSK), 260 ng / ml glial cell growth factor (GGF-2) or 210 ng / ml nerve growth factor (heregulin-betal, HRG)] induced for 10 days, and changed every 72 hours liquid. ...

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Abstract

The invention belongs to the field of cytobiology and relates to a method for efficiently inducing umbilical cord mesenchymal stem cells to differentiate into Schwann-like cells. The method comprises the following steps of: (1) culturing the umbilical cord mesenchymal stem cells to a sub-fusion state, discarding the old culture solution, and adding culture solution comprising 1 mM of beta-mercaptoethanol to induce overnight; (2) discarding the induction solution, washing with a Dulbecco modified eagle medium (DMEM), and adding all-trans retinoic acid to induce for 60 to 84 hours; and (3) discarding the induction solution, washing with the DMEM, adding composition induction solution comprising 5 to 6 ng / ml of platelet-derived growth factors, 9 to 10 ng / ml of alkaline desmocyte growth factors, 13 to 15 uM of Forskolin and 250 to 260 ng / ml of gliocyte growth factors or 200 to 210 ng / ml of nerve growth factors to induce for 10 to 16 days, and change the solution for one time every other 72 hours. By the method, the inductivity can reach over 90 percent.

Description

technical field [0001] The invention belongs to the field of cell biology and relates to a method for inducing cell differentiation. In particular, it relates to a method for rapidly inducing the differentiation of human umbilical cord mesenchymal stem cells into Schwann-like cells. Background technique [0002] With the deepening of tissue engineering artificial nerve research, the research of artificial absorbable materials has gradually matured, but the main bottleneck restricting the development of tissue engineering—the shortage of seed cells is becoming more and more prominent. In the past, Schwann cells were mainly used to construct tissue-engineered artificial nerves. However, Schwann cells are terminally differentiated cells with poor proliferation ability. Adult Schwann cells have even poorer proliferation ability and require pre-denaturation in vitro or in vivo, which is time-consuming, laborious, and difficult. Obtain sufficient Schwann cells in a short period o...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 沈尊理秦金保祝加学
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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