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Method for differentiation of embryonic stem cells (ESC) into mesenchymal stem cells (MSC)

A technology of embryonic stem cells and mesenchymal stem cells, applied in the field of generating mesenchymal stem cells, can solve the problems of long induction time, complicated induction process, and low induction efficiency, and achieve high induction efficiency

Inactive Publication Date: 2015-12-16
ZHEJIANG UNIV
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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for embryonic stem cells to differentiate mesenchymal stem cells, which can solve the shortcomings of the prior art that the induction process is complicated, the induction time is long, and the induction efficiency is not high

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  • Method for differentiation of embryonic stem cells (ESC) into mesenchymal stem cells (MSC)
  • Method for differentiation of embryonic stem cells (ESC) into mesenchymal stem cells (MSC)
  • Method for differentiation of embryonic stem cells (ESC) into mesenchymal stem cells (MSC)

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specific Embodiment approach

[0027] A method for embryonic stem cells to differentiate mesenchymal stem cells, comprising the following steps:

[0028] (1), culture the ESCs until some ESCs tend to differentiate, and obtain fibroblasts;

[0029] (2), ESC in the digestion step (1), the remaining still attached fibrous cells;

[0030] (3), expanding and culturing the remaining attached fibroblasts in step (2);

[0031] (4) Digesting the fibroblasts in step (3), transferring them to an adherent cell culture flask and culturing them to obtain MSCs.

[0032] ESC example picture in step (1) is WA01-Fl (purchased from WiCell company's cell line) cultured on matrigel (purchased from BD company), using mTeSR1 TM 1 (purchased from STEMCELLTECHNOLOGIES company) to maintain culture, such as figure 1 shown. Prolonging the medium exchange time, reducing the frequency of replacing fresh medium mTeSR1 to once every four days, resulting in some ESCs tending to differentiate into fibroblasts, such as figure 2 shown. ...

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Abstract

The invention discloses a method for differentiation of embryonic stem cells (ESC) into mesenchymal stem cells (MSC). The method comprises the following steps of 1, culturing ESC until a part of ESC trends a differentiation state so that fibrous cells are obtained, 2, digesting ESC obtained by the step 1 so that the adhesive fibrous cells are retained, 3, carrying out multiplication culture on the retained fibrous cells obtained by the step 2, and 4, digesting the fibrous cells obtained by the step 3, transferring the digested fibrous cells into an adhesive cell culture bottle and carrying out culture to obtain MSC. The method is free of embryo (EB) culture, realizes direct conversion of ESC into MSC and is simple. Compared with the method comprising direct digestion of ESC by pancreatin, then inoculation and MSC culture, the method provided by the invention has high MSC induction efficiency and short successful induction time.

Description

technical field [0001] The invention relates to a method for generating mesenchymal stem cells (MesenchymalStemCell. MSC) by inducing embryonic stem cells (EmbryonicStemCell, ESC). Background technique [0002] Mesenchymal stem cells (MSCs) are an important member of the stem cell family and a class of pluripotent cells with self-replication and multilineage differentiation capabilities. This type of stem cells can maintain an undifferentiated state and carry out a large number of replication and expansion, and can also undergo multilineage differentiation under certain conditions to form downstream cells of specific tissue types. Most of the differentiated tissue types are tissue types derived from mesoderm, such as bone , cartilage and fat etc. It is derived from the mesoderm and ectoderm in early development and is first found in the bone marrow and later in tissues such as the umbilical cord, placenta, fat and muscle. Current studies have shown that MSCs not only have ...

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 柳华王亚菲蒋诗漫欧阳阳
Owner ZHEJIANG UNIV
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