Characterization and induction of human pre-adipocytes

Human processed lipoaspirate is a source of multipotent adult stem cells [Strem, B.M., Hicok, K.C., Zhu, M., Wulur, L, Alfonso, Z., Schreiber, R.E., Fraser, J.K., Hedrick, M.H., 2005. Multipotential differentiation of adipose tissue-derived stem cells. Keio, Journal of Medicine 54 (3), 132 141] that are able to differentiate between mesenchymal and neurogenic lineage. We characterized PLA cells by cytometry and then they were cultured to induce differentiation into myogenic and neurogenic lineage. Lipoaspirates were digested with collagenase to obtain the pellet, which was labelled with anti-CD44, anti-CD45, and anti-CD90. We used 1313 FACS Calibur flow cytometer to acquire cellular events. Some cells were cultured at 37 degrees C and 5% CO2 in neurogenic or myogenic medium and analysed by immunocytochemistry, using Neuron specific enolase, Vimentin, Glial fibrillary acidic protein, Tau, MAP2 to confirm neurogenic differentiation, MyoD1, Myosin heavy chain, Actin smooth muscle, vimentin to confirm myogenic differentiation. The cytometry results suggest that a part of the cells are of a mesenchymal origin, among which there are progenitor endothelial cells and leucocytes. Microscopy observation already reveals neuronal morphology and longitudinal multinucleated cells compared to control cells. Neurogenic cells only express NSE (early neuronal progenitor marker), but not GFAP, Tau, MAP2 (mature neuron and glial markers); myogenic cells are positive for MyoD1 and Myosin heavy chain. This demonstrates that lipoaspirate cells are capable of differentiating in vitro over a short period of time, and could be employed in biological and clinical research, like mesenchymal adult stem cells. (c) 2006 Elsevier Ltd. All rights reserved.