Effects of Epidermal Growth Factor, Glial Cell Line-Derived Neurotrophic and Leukemia Inhibitory Factor on the Proliferation and Differentiation Potential of Adipose Tissue-Derived Mesenchymal Stem Cells

[Background]There is a great deal of interest in using adipose tissue-derived mesenchymal stem cells (AT-MSCs) for clinical applications. However, the important limitations of clinical application of stem cells are the small number of cells and their differentiation into undesirable lineage in vitro. To overcome this problem, various growth factors are studied extensively.[Objectives]The current study aimed at using 3 different doses of epidermal growth factor (EGF), glial cell line-derived neurotrophic (GDNF), and leukemia inhibitory factor (LIF) to culture AT-MSCs and evaluating their effects on proliferation, viability, differentiation potential, and maintenance of the stemness state of cells.[Methods]The current experimental study was conducted on 8 - 10 male NMRI (Naval medical research institute) mice provided from research center and experimental animal house of Jundishapur University of Ahvaz, Iran, from September 2016 to April 2017. AT-MSCs were isolated from mice adipose tissue. The cells were cultured with three different doses of EGF, LIF, and GDNF. The morphology and cell proliferation of the AT-MSCs were studied on the days 3, 7, and 11 by an inverted microscope and MTT assay, respectively. To evaluate the stemness state of the cells, Oct4 expression was measured using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Also, differentiation potential of AT-MSCs toward adipogenic and osteogenic lineages was assessed. All tests were done in triplicate.[Results]Proliferation and viability of the AT-MSCs cultured in 10 µg/mL EGF, 5 µg/mL LIF, 5 µg/mL GDNF (b2 group) and 20 µg/mL EGF, 5 µg/mL LIF, 5 µg/mL GDNF (b3 group) increased significantly in the days 7 and 11 (170.27 (13.94), 174.39 (18.85) versus 100 (12.08) P < 0.001 (the day 7) and 152.45 (15.75) P < 0.001, 131.53 (19.17) versus 97.64 (13.43) P < 0.022 (the day 11). And differentiation potential of the cells was sustained, but Oct4 overexpressed in treatment groups on the days 7 and 11.[Conclusions]EGF, LIF, and GDNF enhanced proliferation and viability of the AT-MSCs, but for clinical purposes, the growth factors should be applied cautiously.