In vitro differentiation of human multilineage differentiating stress-enduring (Muse) cells into insulin producing cells

Ali M Fouad; Mahmoud M Gabr; Elsayed K Abdelhady; Mahmoud M Zakaria; Sherry M Khater; Amani M Ismail; Ayman F Refaie

doi:10.1016/j.jgeb.2018.09.003

In vitro differentiation of human multilineage differentiating stress-enduring (Muse) cells into insulin producing cells

J Genet Eng Biotechnol. 2018 Dec;16(2):433-440. doi: 10.1016/j.jgeb.2018.09.003. Epub 2018 Dec 5.

Authors

Ali M Fouad¹, Mahmoud M Gabr¹, Elsayed K Abdelhady², Mahmoud M Zakaria¹, Sherry M Khater¹, Amani M Ismail¹, Ayman F Refaie³

Affiliations

¹ Department of Biotechnology, Urology and Nephrology Center, Mansoura 35516, Egypt.
² Department of Zoology, Faculty of Science, Mansoura University, Mansoura 35516, Egypt.
³ Nephrology Department, Urology and Nephrology Center, Mansoura 35516, Egypt.

Abstract

Mesenchymal stem cells (MSCs) is a heterogeneous population. Muse cells is a rare pluripotent subpopulation within MSCs. This study aims to evaluate the pulirpotency and the ability of Muse cells to generate insulin producing cells (IPCs) after in vitro differentiation protocol compared to the non-Muse cells. Muse cells were isolated by FACSAria III cell sorter from adipose-derived MSCs and were evaluated for its pluripotency. Following in vitro differentiation, IPCs derived from Muse and non-Muse cells were evaluated for insulin production. Muse cells comprised 3.2 ± 0.7% of MSCs, approximately 82% of Muse cells were positive for anti stage-specific embryonic antigen-3 (SSEA-3). Pluripotent markers were highly expressed in Muse versus non-Muse cells. The percentage of generated IPCs by flow cytometric analysis was higher in Muse cells. Under confocal microscopy, Muse cells expressed insulin and c-peptide while it was undetected in non-Muse cells. Our results introduced Muse cells as a new adult pluripotent subpopulation, which is capable to produce higher number of functional IPCs.

Keywords: Adipose derived mesenchymal stem cells; Fluorescence activated cell sorting; Mesenchymal stem cells; Muse cells; Real time-quantitative PCR.