[Experiment of osteogenic differentiation of human adipose stem cells in vitro].
OBJECTIVE
To explore the feasibility of isolation, proliferation and osteogenic potential of human adipose stem cells (hASCs) in vitro.


METHODS
hASCs were isolated from the adipose tissue of liposuction aspirated by means of enzymatic digestion and proliferated in vitro. Morphology and growth kinetics by hemacytometer of primary and continuous cells were observed. Osteogenic differentiation of hASCs cultured in conditional culture medium was evaluated by Alkaline phosphatase (ALP) staining and Von Kossa for mineralization, immunohistochemistry of osteocalcin (OCN), ostseopotin (OPN) and collagen type I for matrix maturation, which was further confirmed by RT-PCR analysis for markers of osteogenic differentiation, with the non-osteogenic induced cells as controls. RESUITS: It was shown that both primary and passage cells shared similar fibroblastic-like morphology, while passage cells grew faster. In osteogenic differentiation bioassay, there was a significant difference in expression of ALP, OPN, and OCN between groups when the culture time was extended to 14 days, and collagen type I was expressed in both groups.


CONCLUSIONS
These results indicate that the hASCs grow well in vitro and could be induced to osteoblastic differentiate. hASCs can serve as a seeding cell source for bone engineering.
Wan
Guang-yong
GY
Department of Plastic Surgery, Second Hospital of Shangdong University, Jinan 250033, China. wgykq@163.com
Cai
Jing-long
JL
Zhang
Ming-bin
MB
D000273:Adipose Tissue; D001842:Bone and Bones; D002454:Cell Differentiation; D006801:Humans; D066298:In Vitro Techniques; D010012:Osteogenesis; D013234:Stem Cells; D023822:Tissue Engineering