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A method for promoting cartilage tissue differentiation of mesenchymal stem cells

A technology of cartilage tissue and mesenchymal stem cells, applied in the fields of biology and tissue engineering, to achieve the effect of strong repeatability, easy operation, and promotion of cartilage tissue differentiation of mesenchymal stem cells

Inactive Publication Date: 2019-06-18
杭州枫霖科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the excessive proliferation of mesenchymal stem cells in the process of chondrogenic differentiation is still an insurmountable problem

Method used

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  • A method for promoting cartilage tissue differentiation of mesenchymal stem cells
  • A method for promoting cartilage tissue differentiation of mesenchymal stem cells
  • A method for promoting cartilage tissue differentiation of mesenchymal stem cells

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Embodiment Construction

[0019] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification.

[0020] A method for promoting cartilage differentiation of mesenchymal stem cells, including the following steps: a) isolating human mesenchymal stem cells, b) using lentivirus to stably overexpress NR2F2 gene in mesenchymal stem cells and down-regulating NR2F2 gene expression by small RNA interference technology , C) Culture of 3D cell microspheres after overexpression of NR2F2 gene in mesenchymal stem cells and down-regulation of expression, d) Combining human mesenchymal stem cells, polyethylene glycol dimethacrylate, acryloyl peptide and 2959 photoinitiator And PBS was mixed to prepare bio-ink, e) Use a bioprinter to print the 3D hydrogel cartilage tissue overexpressing the NR2F2 gene on bioprinting paper, f) Culture the 3D hydrogel ...

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Abstract

The invention discloses a method for promoting cartilage tissue differentiation of mesenchymal stem cells. The method comprises the following steps: separating the human mesenchymal stem cells; then, implementing stable over-expression of NR2F2 genes in the mesenchymal stem cells by virtue of lentivirus and down-regulating the expression of the NR2F2 genes by virtue of a small RNA interference technology; cultivating 3D cell microspheres after the over-expression of the NR2F2 genes in the mesenchymal stem cells and down-regulating the expression; mixing the human mesenchymal stem cells, polyethylene glycol dimethacrylate, acrylate peptide, a 2959 photoinitiator and PBS (phosphate buffered saline), so that biological ink is prepared; printing 3D hydrogel cartilage tissues, which achieve the over-expression of the NR2F2 genes, on biological printing paper by virtue of a biological printer; and cultivating the 3D hydrogel cartilage tissues subcutaneously into a mouse by virtue of a cartilage forming medium for 21d, so that joint cartilage tissues are formed. The method provided by the invention is simple and convenient to operate and is strong in repeatability; and the method can effectively promote the cartilage tissue differentiation of the mesenchymal stem cells, so that beneficial reference is provided for the scientific researches of cartilage tissue engineering and the clinical treatment of cartilage injuries.

Description

Technical field [0001] The invention belongs to the technical field of biology and tissue engineering, and specifically relates to a method for promoting the differentiation of cartilage tissue of mesenchymal stem cells. Background technique [0002] In order to maintain the precise printing resolution of 3D printing, the density of cells seeded in bio-ink should not be too high. However, high-density cells are a necessary condition for tissue engineering mesenchymal stem cells to generate extracellular matrix. Mesenchymal stem cells are precursor cells with multiple functions, capable of cartilage, osteogenic and fat differentiation, and can secrete a variety of cytokines. They play an important regulatory role in apoptosis, differentiation, and immune response. Mesenchymal stem cells are easy to separate and collect, and are widely used in the bioprinting process. However, the excessive proliferation of mesenchymal stem cells in the process of cartilage differentiation is stil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/52A61L27/38C12N5/077C12N5/0775C12N5/10
CPCA61L27/3834A61L27/3852A61L27/52A61L2430/06C07K14/70567C12N5/0655C12N2506/1353C12N2513/00
Inventor 崔晓峰高桂芳
Owner 杭州枫霖科技有限公司
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