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Method for producing mesenchymal stem cells through induction of human pluripotent stem cells (hPSC)

A technology of human pluripotent stem cells and mesenchymal stem cells, applied in the field of human pluripotent stem cells to induce mesenchymal stem cells, can solve the problems of laborious differentiation process, low adipogenic differentiation ability, long time, etc., and achieve simple operation and low cost Effect

Pending Publication Date: 2018-11-06
深圳丹伦基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although several methods currently exist to convert hPSCs to hMSCs, each of these methods has its own technical limitations
For example, 1) Some methods of transforming hPSCs into hMSCs involve mouse trophoblast culture system. This differentiation method has the possibility of contamination of mouse trophoblast cells, and has potential risks for later cell transplantation
2) The method involving embryoid body intermediate state formation and flow cytometric sorting of positive cell populations to generate hPSC-MSCs takes a long time and the differentiation process is laborious
3) hPSC-MSCs produced by other differentiation methods show lower adipogenic differentiation ability
4) The use of additional inhibitory factors and special culture systems during the differentiation of hPSCs into hMSCs will increase the production cost of hMSCs

Method used

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  • Method for producing mesenchymal stem cells through induction of human pluripotent stem cells (hPSC)
  • Method for producing mesenchymal stem cells through induction of human pluripotent stem cells (hPSC)
  • Method for producing mesenchymal stem cells through induction of human pluripotent stem cells (hPSC)

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Human induced pluripotent stem cells (hiPSCs) used in this example were induced and differentiated from normal adult male skin fibroblasts.

[0034] 1) HiPSCs were inoculated in Vitronectin (Vitronectin XF)-coated 6-well plates, cultured in TeSR-E8 medium, and replaced with fresh TeSR-E8 medium every other day to continue the culture;

[0035] 2) When the hiPSCs in the 6-well plate reach 60%-70% confluence, wash the cells with PBS and replace the TeSR-E8 medium with the hMSC low glucose medium by one-step method (the one-step method refers to the Directly replace the original medium TeSR-E8 medium with hMSC low-glucose medium), and then replace the fresh hMSC low-glucose medium every 2 days. After 2 weeks of continuous culture, intermediate cell populations gradually formed (see attached figure 1 The microscopic picture of day14 in the medium is shown);

[0036] The hMSC low-glucose medium includes low-glucose DMEM, 10% New Zealand FBS, 1% double antibody (penicilin-st...

Embodiment 2

[0040] Human embryonic stem cells (hESCs) used in this example were induced to differentiate from normal adult male skin fibroblasts.

[0041] The steps for hESC induction to generate MSCs are basically the same as the steps for hiPSC induction in Example 1.

Embodiment 3

[0043] hiPSC-MSC pluripotency differentiation test (fifth generation hiPSC-MSC)

[0044] (1) Identification of osteogenic differentiation

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PUM

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Abstract

The invention provides a method for producing mesenchymal stem cells through induction of human pluripotent stem cells (hPSC). The method comprises the following steps: performing continuous treatmenton the hPSC by a low sugar medium on the basis of a PSC culture system without mouse trophoblast, coating the culture system by gelatin, performing pressure screening on the cells, and performing continuous unicellular passage on the cells through trypsin, and successfully optimizing a simple and high-efficiency method for differentiating the hPSC ino hMSC. The method disclosed by the invention can be used for simultaneously producing lots of functional hPSC-derived hMSC. Meanwhile, the differentiation process is less in time, participation of trophoblast cells, embryoid, flow cytometry, small molecule compounds and special culture conditions is avoided, the operation is simple, and the cost is low.

Description

technical field [0001] The invention relates to the technical fields of biology and medicine, in particular, the invention relates to a method for inducing human pluripotent stem cells to produce mesenchymal stem cells. Background technique [0002] Human pluripotent stem cells (hPSCs), including human induced pluripotent stem cells (hiPSCs) and human embryonic stem cells (hESCs), can serve as an effective cell source for human mesenchymal stem cells (hMSCs). Human mesenchymal stem cells (hMSCs) are a type of adult stem cells derived from human post-embryonic tissues. Because of their self-renewal ability, multilineage differentiation potential, and low oncogenic ability, hMSCs have become an important cell source for tissue engineering, cell therapy, and regenerative medicine. hMSCs have been successfully used clinically in the treatment of a variety of diseases and injuries, including osteogenesis imperfecta, articular cartilage defects, ischemic cardiomyopathy, and nervo...

Claims

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Application Information

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IPC IPC(8): C12N5/0775C12N5/071
CPCC12N5/0662C12N2506/02C12N2506/45C12N2533/52C12N2533/54
Inventor 周焱方崇洲周华涛周海莫翠萍周光前
Owner 深圳丹伦基因科技有限公司
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