Multipotential stem cell-derived mesoderm pedigree mesenchymal stem cell and preparation method thereof

A technology of pluripotent stem cells and mesenchymal stem cells, applied in the field of mesenchymal stem cells of mesoderm lineage and its preparation, can solve the problems of poor treatment effect, cell heterogeneity, uncontrolled differentiation path of cells, etc., and achieve strong proliferation and immune regulation capacity, resolution of source constraints, enhanced cytological or therapeutic effect

Active Publication Date: 2018-04-20
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, the cells obtained by the methods reported in these literatures or patents do not control the differentiation path, only focus on solving the problem of the source of MSCs, and do not provide a corresponding solution for the heterogeneity of the cells, so the induced cells may be better than those derived from bone marrow. Mesenchyme is more heterogeneous; at the same time, these solutions do not focus on the function of cells, and products with poor therapeutic effects will greatly limit their application

Method used

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  • Multipotential stem cell-derived mesoderm pedigree mesenchymal stem cell and preparation method thereof
  • Multipotential stem cell-derived mesoderm pedigree mesenchymal stem cell and preparation method thereof
  • Multipotential stem cell-derived mesoderm pedigree mesenchymal stem cell and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Induction of iPS (induced pluripotent stem cells) to mesoderm mesenchymal stem cells

[0041] Include the following steps:

[0042] 1. Culture of iPS cells

[0043] iPS cells can be conveniently obtained through commercialization or laboratory construction. The present invention focuses on the further cultivation and differentiation operations on the basis of stably established pluripotent stem cell lines. iPS cells can be expanded on a large scale without feeder layers and remain undifferentiated. The culture medium can be selected from the mTeSR series products of STEMCELL company, E8, etc. If it is compatible with the above-mentioned culture medium, it is necessary to coat the culture medium with extracellular matrix, such as Matrigel or laminin. Ensuring high-quality iPS cells is the key to further induction.

[0044] Based on the mTeSR1 and Matrigel culture system, the specific culture operations are as follows:

[0045] 1 Coating of petri dish: Thaw...

Embodiment 2H1

[0068] Example 2 Induction of H1ES embryonic stem cells to mesoderm mesenchymal stem cells

[0069] Include the following steps:

[0070] 1. Culture of H1ES cells

[0071] H1ES cells are readily available commercially. The present invention focuses on the further cultivation and differentiation operations on the basis of stably culturing the H1ES cell line.

[0072] H1ES cells can be expanded on a large scale without feeder layers and remain undifferentiated. The culture medium can be selected from the mTeSR series products of STEMCELL company, E8, etc. If it is compatible with the above-mentioned culture medium, it is necessary to coat the culture medium with extracellular matrix, such as Matrigel or laminin. Ensuring high-quality pluripotent stem cells is the key to further induction.

[0073] Based on the mTeSR1 and Matrigel culture system, the specific culture operations are as follows:

[0074] 1 Coating of petri dish: Thaw Matrigel on ice, dilute Matrigel to 1:20 to...

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Abstract

The invention discloses a multipotential stem cell-derived mesoderm pedigree mesenchymal stem cell and an induced differentiation method thereof. The multipotential stem cell-derived mesoderm pedigreemesenchymal stem cell is prepared by the following steps: performing in-vitro adherent culture on multipotential stem cells and maintaining the undifferentiation state of the multipotential stem cells; preparing unicellular or cell mass suspension from the cells, inoculating to a culture dish coated with matrix glue, and performing cultivation by using the multipotential stem cell culture liquid;after the cells adhere to the wall, adding GSK-3 pathway inhibitor combination into the culture liquid; after growing for 2 to 10 days, obtaining a mesoderm progenitor population; and after subculturing continuously for 2 to 6 times by using mesenchymal stem cell culture liquid, detecting the cell phenotype of mesenchyme. The defects that the human-derived mesenchymal stem cells and the mesenchymal stem cells derived from other non-finite induced way multipotential stem cells have heterogeneity and hybridity are overcome, and the obtained mesoderm pedigree mesenchymal stem cells have higher proliferation capability and immunoregulation capability; and the standardized induced differentiation process can guarantee that the cell populations obtained from different batches have high consistency.

Description

technical field [0001] The invention belongs to the field of biology and medicine, and more specifically, the invention relates to a mesoderm lineage mesenchymal stem cell derived from pluripotent stem cells and a preparation method thereof, including using special culture conditions and selection techniques. The present invention also relates to a therapeutic method using this population of mesenchymal stem cells of mesoderm lineage or its clinical application. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of adult stem cells. They are a large class of pluripotent cells that exist in many tissues in the adult body, such as bone marrow, umbilical cord blood, fat, and peripheral blood. They are in a quiescent state under normal conditions. When the tissue or organ where they are located is damaged, they can quickly enter a proliferative state and maintain a great proliferative potential, and differentiate into different cell types to repair the damage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2500/30
Inventor 项鹏李伟强翟志臣宋武
Owner SUN YAT SEN UNIV
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