Isolation, culture and identification of human adipose-derived mesenchymal stem cells

Lin chuang er bi yan hou tou jing wai ke za zhi = Journal of clinical otorhinolaryngology, head, and neck surgery
W J Zhang, J J Sun

Abstract

Objective: To establish isolation and culture methods of human adipose derived mesenchymal stem cells(hADSCs) in vitro and to investigate the basic biological characteristics of hADSCs as seed cells involved in repair of epithelium during temporal bone surgery.Method:Human adipose tissue was collected after liposuction surgery from the cosmetic and plastic surgery center,and then was digested with 0.3% collagenase Ⅰ. hADSCs were isolated and adherent cultured. Multilineage differentiation potential of hADSCs were determined by differentiating cells into adipocytes and osteoblast. Phenotypes of cells were detected by flow cytometer, and cell growth curve was observed by MTT. Result:Obtained hADSCs were similar size, spindle-shaped fibroblasts-liked cells and had differentiation potential to adipocytes and osteoblast. Flow cytometry showed that cells were highly positive for CD105, CD90, CD29 and weakly positive for CD34 and CD45. MTT exhibited that hADSCs had a exponential growth phase. Conclusion:Isolated stem cells from human adipose tissue show the typical characterization of hADSCs, and can be used as seed cells involved in repair of epithelium during temporal bone surgery.

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