Method for improving osteogenesis capacity of mesenchymal stem cells

A technology of stem cells and capacity, applied in animal cells, vertebrate cells, cell culture active agents, etc., can solve problems such as low efficiency

Pending Publication Date: 2020-10-30
山东佰傲干细胞生物技术有限公司
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Problems solved by technology

At present, promoting the differentiation of mesenchymal stem cells into osteoblasts and improving the effect of bone regeneration and repair is an important issue in scientific research and clinical application in relat

Method used

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  • Method for improving osteogenesis capacity of mesenchymal stem cells
  • Method for improving osteogenesis capacity of mesenchymal stem cells
  • Method for improving osteogenesis capacity of mesenchymal stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0044] 1. Isolation and culture of human umbilical cord mesenchymal stem cells

[0045] 1. In a biosafety cabinet, the surface of the umbilical cord was fully rinsed with phosphate buffered solution (PBS), the superficial membrane of the umbilical cord tissue was peeled off, and then the umbilical cord was cut open to remove the residual blood in two umbilical arteries and one umbilical vein. And remove arteriovenous vessels. Cut the processed umbilical cord tissue into a size of about 1-2mm 3 Then place the tissue block in α-MEM medium containing 10% FBS, shake the tissue block and spread it on the culture plane, then put it in the medium containing 5% CO 2, Cultivated in a saturated humidity incubator at 37°C, the frequency of changing the culture medium is once every two days, pay close attention to the growth status of the cells, and the time of changing the medium can be adjusted according to the growth of the cells.

[0046] 2. Observe that the cells cover about 80% ...

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Abstract

The invention provides a method for improving the osteogenesis ability of mesenchymal stem cells, which comprises the following steps: inoculating the mesenchymal stem cells into an alpha-MEM culturemedium containing 10% of fetal calf serum, culturing, digesting and subculturing by 0.2% of pancreatin when the fusion degree of the mesenchymal stem cells is 80-85%, inoculating the obtained mesenchymal stem cells into an alpha-MEM culture medium containing 10% of fetal calf serum for culture, and adding osteogenesis induction liquid and a metformin aqueous solution for induction when the densityof the mesenchymal stem cells is greater than 90% for induction for 14-21 days. The metformin acts on the mesenchymal stem cells, alizarin red staining results show that the metformin can significantly improve the calcium mineralization nodule degree of the cells, and it is indicated that the metformin has a significant induction effect on mesenchymal stem cell osteogenic differentiation, the metformin can significantly improve the expression of the mesenchymal stem cell osteogenic differentiation related genes SIRT1, ALP, COL1A1, RUNX2 and BGLAP, and the improvement of the osteogenic differentiation capacity of the mesenchymal stem cells is of great significance to stem cell transplantation and bone biology research.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a method for improving the osteogenic ability of mesenchymal stem cells. Background technique [0002] Bone tissue engineering refers to the cultivation and expansion of isolated autologous high-concentration osteoblasts, bone marrow stromal stem cells or chondrocytes in a natural or synthetic biocompatibility that can be gradually degraded by the human body. On the absorbed cell scaffold or extracellular matrix, this kind of biomaterial scaffold can provide cells with a three-dimensional living space, which is beneficial for cells to obtain sufficient nutrients, perform gas exchange, and remove waste materials, so that cells can be placed on the prefabricated three-dimensional scaffold. Then, the cell hybrid material is implanted into the bone defect site. While the biomaterial is gradually degraded, the implanted bone cells continue to proliferate, so as to achieve the purpos...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0654C12N2506/1353C12N2506/1384C12N2506/1369C12N2506/1392C12N2500/38C12N2501/999C12N2501/39C12N2500/42
Inventor 高迎凤蒲锋星程蕊苹
Owner 山东佰傲干细胞生物技术有限公司
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