Authors: Lubos Danisovic, Bohac Martin, Stefan Polak, Stanislav Ziaran, Lenka Oravcova and Marcela Kuniakova
The purpose of present study was isolation and in vitro expansion of human adipose tissue-derived stem cells ATSCs) and analysis of long-term cultivation effect on their biological properties. Adipose tissue was obtained by liposuction from healthy individuals during planed aesthetic surgeries. ATSCs were isolated by collagen type I and were cultured up to 30th passage. The morphology was analyzed by inverted microscope. During passages the kinetics of ATSCs proliferation was evaluated by growth curves. Cellular senescence was assessed by β-Galactosidase staining kit. Moreover, the chondrogenic potential was analyzed by real time PCR and the cytogenetic stability was assessed as well. Obtained results showed: i) alterations in shape of ATSCs in increasing passages; ii) minor decreasing of their proliferation capacity; iii) stable normal count of chromosomes; iv) about 30% of ATSCs undergo replicative senescence; v) unchanged chondrogenic potential. According these findings it should be emphasize that long-term cultivation (up to 30th passage) do not seriously affect biological properties of human ATSCs.