Trans differentiating human adipose-derived mesenchymal stem cells into male germ-like cells utilizing Rabbit Sertoli...

Mesenchymal stem cells (MSCs) are deemed as potential new therapeutic agents for infertility treatment and adipose tissue (AT) becomes a potential MSCs source. To direct MSCs through the differentiation process properly, an environment comparable to the in vivo niche might be indispensable.

This study aims to differentiate human AT-derived MScs (hAD-MScs) into male germ-like cells in vitro using a combination of rabbit Sertoli cells conditioned medium (SCCM), bone morphogenetic protein 4, and retinoic acid.

MScs were isolated from human ATs of fertile and infertile donors. The verified MScs were differentiated using a 2-step protocol; the first step included 20 ng/ml bone morphogenetic protein 4 treatment. The second step was performed utilizing 1 μM retinoic acid and/or SCCM. The morphological changes and the expression of germ cell (GC)-specific markers: octamer-binding transcription factor-4; stimulated by retinoic-acid-8, synaptonemal complex protein-3, andprotamine-1 were assessed in the treated cells using quantitative polymerase chain reaction.

Induction of hAD-MScs resulted in the upregulation of GC-specific genes where SCCM treatment showed the highest expression. The synaptonemal complex protein-3andprotamine-1 gene expression was detected after 19 and 26 days of induction, respectively. PRM1 was detected in hAD-MScs cultured in SCCM earlier than in other treated groups. The treated cells became more elongated-like spindles and formed aggregates.

hAD-MScs differentiated to GC lineage exhibited the ability to express GC-specific markers under in vitro conditions, and rabbit's Sertoli cells can be used for inducing transdifferentiation of hAD-MScs into germ-like cells.