Alaa Mutee' Khudair - Medical Laboratory Sciences Department, Faculty of Health Sciences, Islamic University of Gaza, Gaza City, Palestine.
Mazen Medhat Alzaharna - Medical Laboratory Sciences Department, Faculty of Health Sciences, Islamic University of Gaza, Gaza City, Palestine.
Fadel Akram Sharif - Medical Laboratory Sciences Department, Faculty of Health Sciences, Islamic University of Gaza, Gaza City, Palestine.

Background: 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. Objective: 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 ۴, and retinoic acid. Materials and Methods: MScs were isolated from human ATs of fertile and infertile donors. The verified MScs were differentiated using a ۲-step protocol; the first step included ۲۰ ng/ml bone morphogenetic protein ۴ treatment. The second step was performed utilizing ۱ μM retinoic acid and/or SCCM. The morphological changes and the expression of germ cell (GC)-specific markers: octamer-binding transcription factor-۴; stimulated by retinoic-acid-۸, synaptonemal complex protein-۳, and protamine-۱ were assessed in the treated cells using quantitative polymerase chain reaction. Results: Induction of hAD-MScs resulted in the upregulation of GC-specific genes where SCCM treatment showed the highest expression. The synaptonemal complex protein-۳ and protamine-۱ gene expression was detected after ۱۹ and ۲۶ days of induction, respectively. PRM۱ 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. Conclusion: 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.

Adipose tissue-derived mesenchymal stem cell, Bone morphogenetic protein ۴, Germ-line cells, Retinoic acid, Sertoli cells., سلول های بنیادی مزانشیمی مشتق از بافت چربی, پروتئین مورفوژنتیک استخوان ۴, سلول های زایا, رتینوئیک اسید, سلول های سرتولی.