Isolation of adipose stem cells (ASCs) subpopulations with distinct differentiation potential

cultured with Basal Medium until the confluence, then the cells were trypsinized and divided in 3 groups. The first one was used to per- form RT Real-Time PCR for CD44, CD105; CD73, CD90 and STRO-1. The second group was cultured with Osteogenic Medium for 3 weeks. The third one was used to set up a pellet culture with chondrogenic medium and cultured for 3 weeks. After 3 weeks of culture with osteogenic medium or chondrogenic medium the sam- ples were retrieved. Alizarin Red Staining and RT Real-Time PCR for Osteocalcin and Osteopontin were used to establish the osteo- blast differentiation potential. Alcian Blue, Toluine Blue and Sa- franin O staining and RT Real-Time PCR for Agrecan, Collagen I, Collagen II, Collagen X and Sox 9 assess the chondrogenic differ- entiation potential. The described method was effective to isolate distinct subpopulations which present different gene expression profile relative to the stem cell markers studied. The expression of osteogenic and chondrogenic markers showed that these subpopu- lations exhibit significantly different differentiation potentials.

Acknowledgements: Marie Curie Actions Alea Jacta Est, Project HIPPOCRATES, NoE EXPERTISSUES.