Method for enhancing mesenchymal stem cell chemotactic capacity and chemotactic factor CCL5 expression

A technology of mesenchymal stem cells and chemokines, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of limited isolation of CCL5, difficulty in meeting the needs of biological function research and clinical application, and achieve improvement The effect of migratory ability

Active Publication Date: 2016-02-17
XIAMEN UNIV
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Problems solved by technology

However, the isolation of CCL5 from natural tissues is limited, and it is difficult to meet the needs of its biological function research and clinical application, so increasing the expression of CCL5 is conducive to in-depth basic and clinical research on CCL5

Method used

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  • Method for enhancing mesenchymal stem cell chemotactic capacity and chemotactic factor CCL5 expression
  • Method for enhancing mesenchymal stem cell chemotactic capacity and chemotactic factor CCL5 expression
  • Method for enhancing mesenchymal stem cell chemotactic capacity and chemotactic factor CCL5 expression

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Embodiment Construction

[0055] The following embodiments will further illustrate the present invention in conjunction with the accompanying drawings.

[0056] (1) Isolation and primary culture of umbilical cord mesenchymal stem cells

[0057] Take the baby's umbilical cord tissue, soak it in MSCs medium MSCM (stemcell company), wash the residual blood with PBS in an ultra-clean bench, remove the umbilical vein, umbilical artery and umbilical cord adventitia, peel off Wharton's glue, and cut into 1-2mm pieces 3 The size of the tissue block; transfer the tissue block to 0.1% type Ⅱ collagenase, add a little MSCM, digest overnight in a constant temperature shaker at 37°C until the Wharton glue is completely digested; suck the digestion solution containing cells into a sterile centrifuge tube , blow the cells with 30 times the volume of MSCM, centrifuge at 2000r / min for 10min, discard the supernatant; resuspend the cells in -MEM medium supplemented with 10% fetal bovine serum, inoculate into a culture bo...

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Abstract

The invention discloses a method for enhancing mesenchymal stem cell chemotactic capacity and chemotactic factor CCL5 expression, and relates to mesenchymal stem cells. The method comprises the steps of separation of the mesenchymal stem cells and passage of primary culture umbilical cord mesenchymal stem cells; identification of the umbilical cord mesenchymal stem cells; stimulation of TNF-alpha and IFN-gamma to hUC-MSCs; detection of CCL5 expression through Real-Time PCR; detection of CCL5 expression through ELISA. CCL5 expression through MSCs is improved by 924 times, a large amount of natural CCL5 can be obtained, no lipofection transfection is needed, and rapidness and convenience are achieved. Due to simulation of TNF-alpha and IFN-gamma, the transfer ability of the mesenchymal stem cells is improved, it is promoted that the mesenchymal stem cells can more rapidly collect tissue injuries and inflammation portions in vivo, and the functions of tissue repairing and immune modulating are brought into play.

Description

technical field [0001] The invention relates to mesenchymal stem cells, in particular to a method for enhancing the chemotactic ability of mesenchymal stem cells and the expression of chemokine CCL5. Background technique [0002] Mesenchymal stem cells (mesenchymal stem cells) are a type of adult stem cells with multi-directional differentiation potential, have immune regulation, damage repair, can secrete a variety of active factors, target damage, tumors and other parts; it is the research and treatment of tissue damage, immune suppression ideal materials and tools for diseases such as [0003] Chemokine refers to a class of small molecular proteins that have chemotactic effects on inflammatory cells. According to the distribution of cysteine ​​residues at the four sites in its molecular structure and the structure of nearby amino acid residues, are CXC, CC, C and CX3C4 species (Balkwill FR. The Journal of Pathology, 2012; 226(2): 148-157). The chemokine CCL5 is a member...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12Q1/68G01N33/68
Inventor 宋刚钟伟
Owner XIAMEN UNIV
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