Human adipose tissue provides abundant source of mesenchymal stem cells, which can be easily and safely harvested as compared with human bone marrow.
The aim of this study was to differentiate a viable osteoblast and adipocyte cell culture from mesenchymal stem cells, an experimental model to study the interrelation between adipocytes and bone forming cells, osteoblasts.
Materials and methods: Subcutaneous adipose tissue liposuction aspirate was obtained from a female subject, aged 27 years undergoing plastic surgical procedure in a specialized clinic, after she gave the informed consent.
The adipose tissue sample was extensively washed with sterile PBS containing antibiotic/antimycotic (AA) and let 30 min to decant. Blood fraction was collected into 50 ml conical tubes. Cells were cultured in DMEM, 10% FBS, AA. After 3 weeks, cells were splitted and seeded in osteogenic (DMEM, 10% FBS, 100 nm dexametasone, 10 mm beta glycerolphosphate, 0.05 mm ascorbic acid, 1% AA) and adipogenic medium (DMEM, 10% FBS, 1 μm dexametasone, 10 μm insulin, 200 μm indomethacin, 0.5 μm IBMX, 1% AA) respectively. Specific phenotype was assessed by von Kossa specific staining for osteoblasts and Oil red staining for adipocytes.
Results: The specific phenotype occurred in the second passage. Cell growth rate was higher in osteoblasts than adipocytes (4.5 vs 3.2 in passage 7). Osteoblasts were fully diferentiated (mineralized) in passage six. The log phase of the cellular growth curve was 10 passages long, that offers a reasonable period of time for experiments that require adipocyte-osteoblast co/culture.
Conclusion: Differentiation of osteoblasts and adipocytes starting from the same cellular sample from human adipose-tissue stromal derived cells confers a great advantage for co-culture studies because immunological rejection is overcome.
Declaration of interest: The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project.
Funding: This work was supported, however funding details unavailable.
05 - 09 May 2012
European Society of Endocrinology