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[Isolation, culturation and adipogenisis committed differentiation of adult human mesenchymal stem cell]
Hang-hui Cen1, Chun-mao Han, Ping-ping Lai
1The Second Affiliated Hospital, College of Medical Sciences, Zhejiang University, Hangzhou 310009, China.
Objective:
To isolate MSCs from adult human bone marrow cells and to induce them into adipocytes.
Methods:
MSCs were isolated from adult human bone marrow aspirated by Percoll and expanded in L-DMEM. The surface antigen of MSCs, CD14, CD34, CD45, CD44, VLA-1, HLA-DR and cell cycle were analysed on a FACScan flow cytometer. MSCs were cultured in adipogenisis inducing medium including insulin, 1-methyl-3-isobutylxanthine, indomethine and dexamethasone for 7 days and stained with Oil Red O.
Results:
MSCs grew as adherent cells and expanded more than 10 passages. They were positive for CD44 and negative for CD14, CD34, CD45, HLA-DR. The expression of VLA-1 was weak. After 7 days of adipocyte inducing, about 85%of the cells displayed accumulation of lipid vacuoles, as detected by Red Oil O.
Conclusion:
MSCs isolated and cultured from adult human bone marrow can be induced to adipogenisis committed differentiation.
Insights
Researchers isolated mesenchymal stem cells (MSCs) from human bone marrow and successfully induced them into adipocytes, demonstrating their potential for differentiation.
Area of Science:
- Stem Cell Biology
- Cell Differentiation
- Regenerative Medicine
Background:
- Mesenchymal stem cells (MSCs) are multipotent stromal cells with the capacity for differentiation into various cell types.
- Bone marrow is a rich source of MSCs for research and therapeutic applications.
- Adipogenesis, the process of fat cell formation, is a key differentiation pathway for MSCs.
Purpose of the Study:
- To isolate mesenchymal stem cells (MSCs) from adult human bone marrow.
- To induce the isolated MSCs into adipocytes (fat cells).
- To characterize the differentiation potential of bone marrow-derived MSCs.
Main Methods:
- MSCs were isolated from adult human bone marrow using Percoll density gradient centrifugation.
- Cell surface markers (CD14, CD34, CD45, CD44, VLA-1, HLA-DR) were analyzed by flow cytometry.
- Adipogenic differentiation was induced using a specific medium containing insulin, IBMX, indomethacin, and dexamethasone for 7 days.
- Lipid accumulation was visualized using Oil Red O staining.
Main Results:
- Isolated MSCs exhibited adherence and expanded beyond 10 passages.
- Flow cytometry confirmed MSCs were positive for CD44 and negative for CD14, CD34, CD45, and HLA-DR, with weak VLA-1 expression.
- Approximately 85% of the differentiated cells showed significant lipid vacuole accumulation, confirmed by Oil Red O staining.
- The results indicate successful induction of adipogenesis in cultured MSCs.
Conclusions:
- Mesenchymal stem cells isolated and cultured from adult human bone marrow possess the capacity for adipogenic differentiation.
- These findings support the use of bone marrow-derived MSCs for studying adipogenesis and potentially for therapeutic applications involving fat tissue regeneration.