1. Emu oil Stabilized Stemness in Adipose Tissue derived Stem Cells  Khatereh Saei Arezoumand1*, Effat Alizadeh1, 2, Mohammad Esmaeilluo3, Yones Pilehvar-Soltanahmadi1,2, and Nosratollah Zarghami1, 2* 1Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran 2The Umbilical Cord Stem Cell Research Center (UCSRC), Tabriz University of Medical Sciences, Tabriz, Iran 3Universita degli studi di siena Department of Medical Biotechnologies Siena, Italy  
Introduction:
Recent evidence suggests that adipose tissue derived mesenchymal stem cells (AT-MSCs) have promising therapeutic potential for a broad range of diseases. Because the percentage of AT-MSCs obtained from adipose tissue is very low for cell therapy applications, ex vivo expansion of AT-MSCs is necessary, but aging, loss of stemness and undesired differentiation of them during in vitro cultivation reduces their effectiveness. For achieving ideal therapeutic potential of AT-MSCs in tissue regenerative purposes, it is necessary to retain their stemness properties in vitro.
Emu oil contains fatty acids and antioxidants,which showed anti-inflammatory effects and promotes wound healing by accelerating the growth rate of keratinocytes in recent studies.The aim of this study was to investigate the effect of emu oil on MSCs stemness.
Results:
Isolated AT-MSCs in passage 3 were positive for expression of CD markers CD90, CD73, CD105 but negative for CD45. Also, adipogenic and osteogenic potential of AT-MSCs were confirmed. MTT results showed significant higher (p<0.05) proliferation and viability in emu oil treated stem cells as compared to control. The expression of marker genes were significantly higher (p<0.05) in emu oil treated AT-MSCs compared to control. Colony forming assay of AT-MSCs treated with emu oil showed 2 folds higher number of colonies in emu oil treated stem cells.
RNA extraction (Nano drop)
MTT cell growth measurement assay
Material methods:
In this study, the AT-MSCs isolated from subcutaneous adipose tissue, and characterized by FACS.For plasticity evaluation,AT-MSCs induced for adipogenic and osteogenic differentiation. For addition of emu oil in to medium, it was emulsified using lecithin and toluene. The effect of emu oil on viability and proliferation of AT-MSCs was evaluated using MTT-assay. The expression of multipotency marker genes including Nanog, OCT-4, SOX2, Nestin, and Rex1 were investigated on days 1, 3,5 and 9 days after cultivation in the presence of emu oil with QRT-PCR.Colony forming assay was performed for evaluation of the e ffect of emu oil on AT-MSCs colony formation.
Cell Culture
MTT Assay
Real Time PCR results
Colony Forming Assay
Conclusion:
These results suggest that addition of emu oil may act as a supplement to AT-MSCs culture for stabilizing their stemness. The findings of this study could be applicable for improving AT-MSCs quality used in cell therapy.
Emulsifiers
Sonicator