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Method for preparing clinical-grade mesenchymal stem cell preparation by utilizing human induced pluripotent stem cells

A technology of pluripotent stem cells and stem cells, applied in artificially induced pluripotent cells, biochemical equipment and methods, animal cells, etc., can solve the problems of low homogeneity, unstable quality and limited sources, and reach the market The effect of broad prospects, high homogeneity and unlimited sources

Active Publication Date: 2020-12-08
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

[0003] However, due to the deficiencies of the existing technology and clinical needs, mesenchymal stem cells still have limited sources, unstable quality, and low homogeneity in cell transplantation therapy and tissue engineering for articular cartilage damage and degenerative diseases.

Method used

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  • Method for preparing clinical-grade mesenchymal stem cell preparation by utilizing human induced pluripotent stem cells
  • Method for preparing clinical-grade mesenchymal stem cell preparation by utilizing human induced pluripotent stem cells
  • Method for preparing clinical-grade mesenchymal stem cell preparation by utilizing human induced pluripotent stem cells

Examples

Experimental program
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Embodiment 1

[0050] A method for preparing a clinical grade mesenchymal stem cell preparation using human induced pluripotent stem cells, comprising the steps of:

[0051] 1. Conventional culture of human iPSC: the human iPSC cell line was purchased from Nuwacell Company, which is a NuwacellTM scientific research grade hiPSC cell line (RC01001-B), which was inoculated in 10 μg / ml Vitronectin XF TM (composition includes 960μl CellAdhere TM DilutionBuffer+40μl 250μg / ml Vitronectin XF TM ) coated 6-well plate, using iPSC regular medium (1920μl TeSR TM -E8 TM Basal Medium+80μl 25×TeSR TM -E8 TM Supplement) at 37°C, 5% CO 2 Cultivate for 6 days under saturated humidity conditions. The above conventional medium was replaced with fresh one every day to continue culturing. During this period, the morphology of human iPSCs at different culture times was photographed with an optical microscope. see results figure 1. Undifferentiated human iPSCs were grown in 6-well plates on Day-2 before d...

Embodiment 2

[0060] A method for preparing a clinical grade mesenchymal stem cell preparation using human induced pluripotent stem cells, comprising the steps of:

[0061] 1. Routine culture of human iPSC: Human iPSC cell line (purchased from Nuwacell) was inoculated in 10 μg / ml Vitronectin XF TM (Consists of 960μl CellAdhere TM DilutionBuffer+40μl 250μg / mlVitronectin XF TM ) coated 6-well plate, using iPSC regular medium (1920μl TeSR TM -E8 TM Basal Medium+80μl 25×TeSR TM -E8 TM Supplement) at 37°C, 5% CO 2 Cultivate for 6 days under saturated humidity conditions. The above conventional medium was replaced with fresh one every day to continue culturing. During this period, the morphology of human iPSCs at different culture times was photographed with an optical microscope. see results figure 1 . Undifferentiated human iPSCs were grown in 6-well plates on Day-2 before differentiation to a 70% compact cobblestone-like multicellular colony with a high nucleoplasmic ratio and promin...

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Abstract

The invention provides a method for preparing a clinical-grade mesenchymal stem cell preparation from human induced pluripotent stem cells, and belongs to the technical field of cell culture. The method for preparing the clinical-grade mesenchymal stem cell preparation by utilizing the human induced pluripotent stem cells comprises the following steps of induced differentiation of iPSC to iPSC-MSC, amplification of iPSC-MSC and preparation of the iPSC-MSC preparation, and further comprises identification of iPSC-MSC characteristics and quality control of the iPSC-MSC preparation. The clinical-grade mesenchymal stem cell preparation prepared by the invention has the advantages of providing mesenchymal stem cell sources which are infinite in source, stable in quality, high in homogeneity, relatively controllable and the like in response to individual requirements of various patients, realizes large-scale industrialization, and has the characteristic of a broad market prospect regarding its clinical application.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for preparing a clinical-grade mesenchymal stem cell preparation by using human induced pluripotent stem cells. Background technique [0002] Mesenchymal stem / stromal cell (MSC) cultured in organ tissue and in vitro has natural high heterogeneity, which is not only the basis for its multiple functional activities, but also affects the repeatability of MSC experiments, the quality of MSC and even the clinical efficacy. The limiting factors of these factors have caused more variables and difficulties to standardize and standardize the evaluation of the safety and efficacy of MSC treatment. Human induced pluripotent stem cells (iPSC) and related studies based on cell reprogramming technology have significantly improved the understanding of stem cell maintenance of pluripotency and stemness and their molecular regulation mechanisms, while also providing an u...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28C12Q1/02
CPCC12N5/0662A61K35/28C12N2506/45
Inventor 张娟周光前
Owner 深圳丹伦基因科技有限公司
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