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Culture system beneficial for mesenchymal stem cells to be applied in bone regeneration

A technology of mesenchymal stem cells and culture system, applied in cell culture active agent, culture process, tissue regeneration, etc.

Inactive Publication Date: 2017-05-24
TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Placental and adipose-derived mesenchymal stem cells, like other tissue-derived mesenchymal stem cells, determine key transcription factors (Osterix, RUNX) and early representative factors of bone regeneration (osteopontin-OPN, basic phosphatase-ALP) gene expression was up-regulated, but the process of osteogenic induction required at least 14 days

Method used

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  • Culture system beneficial for mesenchymal stem cells to be applied in bone regeneration
  • Culture system beneficial for mesenchymal stem cells to be applied in bone regeneration
  • Culture system beneficial for mesenchymal stem cells to be applied in bone regeneration

Examples

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Embodiment 1

[0042] A culture system conducive to the application of placental mesenchymal stem cells in bone regeneration, the steps are as follows:

[0043] ⑴Isolation of placental mesenchymal stem cells: obtain sterile placental tissue, peel off the outer membrane of the placental tissue and the internal artery and vein of the placental tissue under aseptic conditions, separate the placental Wharton's jelly and cut it into 2-3mm 3 , paste the shredded Wharton's glue on the bottom of the culture bottle, and add an appropriate amount of culture solution to the culture bottle after 1-4 hours. The culture solution uses a-MEM+5% platelet lysate+50mg / ml ascorbic acid+10nmol / L EGF +10nmol / L bFGF+10nmol / L recombinant human brain natriuretic peptide (hBNP);

[0044] (2) Digestion and passage of placental mesenchymal stem cells. When the placental mesenchymal stem cells grow to 80%, they are digested with 0.05% trypsin + 0.02% EDTA, and 1×10 4 cells / cm 2 Transfer to a new culture bottle;

[00...

Embodiment 2

[0050] A culture system conducive to the application of umbilical cord mesenchymal stem cells to bone regeneration, the steps are as follows:

[0051] (1) Separation of umbilical cord mesenchymal stem cells: obtain sterile umbilical cord tissue, peel off the outer membrane of the umbilical cord tissue and the internal artery and vein of the umbilical cord tissue under aseptic conditions, separate the umbilical cord Wharton's jelly and cut it into 2-3mm 3 , paste the shredded Wharton's glue on the bottom of the culture bottle, and add an appropriate amount of culture solution to the culture bottle after 1-4 hours. The culture solution uses a-MEM+5% platelet lysate+50mg / ml ascorbic acid+10nmol / L EGF +10nmol / L bFGF+10nmol / L recombinant human brain natriuretic peptide (hBNP);

[0052] (2) Digestion and passage of umbilical cord mesenchymal stem cells. When the umbilical cord mesenchymal stem cells grow to 80%, digest them with 0.05% trypsin + 0.02% EDTA, and dilute them with 1×10 ...

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Abstract

The invention relates to a culture system beneficial for mesenchymal stem cells to be applied in bone regeneration, wherein the culture system includes a-MEM+5% platelet lysate+50 mg / ml ascorbic acid +10 nmol / L EGF+10 nmol / L bFGF+10 nmol / L recombinant human brain natriuretic peptide. Placenta-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells cultured by the culture system all can be used as seed cells to be applied in bone tissue engineering treatment of bone defects, the step of mesenchymal stem cell in-vitro osteogenic induction is avoided, and the time for clinical application of bone tissue engineering treatment of bone defects is shortened by at least 14 days. In addition, compared with the adipose-derived mesenchymal stem cells, with application of the cultured placenta-derived mesenchymal stem cells, a bone regeneration factor gene is significantly highly expressed.

Description

technical field [0001] The invention belongs to the field of biotechnology and is a culture system that is beneficial to the application of placental mesenchymal stem cells in bone regeneration. Background technique [0002] Bone tissue defect is caused by trauma, infection, tumor and other factors that cause the body to lose some bone mass and form a large gap. This problem is relatively common in clinical practice and difficult to deal with. How to promote the repair of bone tissue defects has always been the focus of research by researchers in this field. In recent years, with the development of bone tissue engineering technology, this problem has been solved to a certain extent. Seed cells play a key role in bone tissue engineering. Therefore, the selection of seed cells is very important. Early researchers used bone cells and bone marrow stromal cells, but these seed cells are difficult to obtain, which is likely to cause secondary damage to patients, and the ability ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61L27/38A61L27/50
CPCC12N5/0667A61L27/3834A61L27/3847A61L27/50A61L2430/02C12N5/0668C12N2500/30C12N2500/38C12N2501/11C12N2501/115C12N2501/999
Inventor 赵刚马洁刘微微高伟玮
Owner TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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