Detection method and detection reagent for osteogenic potential of mesenchymal stem cells

A technology of mesenchymal stem cells and detection methods, which is applied in the field of detection methods and detection reagents for the osteogenic potential of mesenchymal stem cells, and can solve the problems of excessively increasing culture time and culture costs

Pending Publication Date: 2022-01-04
上海葆年生物科技有限公司
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Problems solved by technology

If the induced differentiation of mesenchymal stem cells fails, the culture time and culture cost will be excessively increased, so it is necessary to provide a detection method and detection reagent that can detect the osteogenic potential of mesenchymal stem cells during the directional induction culture process

Method used

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  • Detection method and detection reagent for osteogenic potential of mesenchymal stem cells
  • Detection method and detection reagent for osteogenic potential of mesenchymal stem cells
  • Detection method and detection reagent for osteogenic potential of mesenchymal stem cells

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specific Embodiment 1

[0057] 1) Resuscitate the mesenchymal stem cells until they reach 70%-90%; 2) Digest and passage the mesenchymal stem cells, and culture them with complete medium for 1-3 days; 3) When the cell confluence reaches 80%-90%, suck away Complete medium, add mesenchymal stem cell osteogenic induction and differentiation medium for culture, and change the medium every 3-4 days; 4) On the 5th, 10th and 15th day of continuous induction, real-time quantitative PCR method was used to detect osteogenic specificity Genes, the target genes include osteocalcin (osteocalcin, OCN) and human RUNX family transcription factor 2 (Homo sapiens RUNX family transcription factor 2, RUNX2), in order to detect osteocalcin, human Expression of RUNX family transcription factor 2.

[0058] 1. Extraction of total RNA from each group of cells

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Abstract

The invention relates to a detection reagent and a detection method for osteogenic potential of mesenchymal stem cells, and the detection method comprises the step of determining the osteogenic potential of mesenchymal stem cells by detecting the expression quantity of osteocalcin and human RUNX family transcription factor 2 in osteogenic induction of mesenchymal stem cells. According to the method for detecting the osteogenic potential of the mesenchymal stem cells, the osteogenic potential of the mesenchymal stem cells is determined by detecting the expression quantity of osteocalcin and a human RUNX family transcription factor 2 in osteogenic induction of the mesenchymal stem cells, the osteogenic potential of the mesenchymal stem cells can be determined through specific gene expression detection in the early and middle stages of induced differentiation, and the osteogenic potential of the to-be-detected mesenchymal stem cells is known, so that the method has important significance on the osteogenic induction culture process of the mesenchymal stem cells.

Description

technical field [0001] The present application relates to the technical field of stem cell osteogenesis induction culture, in particular to a detection method and a detection reagent for the osteogenic potential of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of adult stem cells with high self-renewal ability and multi-directional differentiation potential, widely present in bone marrow, fat, amniotic membrane, placenta, umbilical cord blood and umbilical cord tissue It can be induced to differentiate into various cells and tissues such as bone, fat, cartilage, nerve and cardiac muscle. [0003] Osteogenic induction and differentiation of stem cells is usually carried out in vitro. Due to the complexity of the mechanism of stem cell differentiation, the cost of in vitro directional induction and differentiation is much more expensive than that of traditional ordinary cell culture. In induction culture of mesenchymal stem cel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/158C12Q2600/166
Inventor 崔晓燕
Owner 上海葆年生物科技有限公司
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