TY - JOUR AU - Alaa Mutee’ Khudair AU - Mazen Medhat Alzaharna AU - Fadel Akram Sharif PY - 2023/04/11 Y2 - 2024/03/28 TI - Trans differentiating human adipose-derived mesenchymal stem cells into male germlike cells utilizing Rabbit Sertoli cells: An experimental study JF - International Journal of Reproductive BioMedicine (IJRM) JA - IJRM VL - 21 IS - 3 SE - Articles DO - 10.18502/ijrm.v21i3.13197 UR - https://knepublishing.com/index.php/ijrm/article/view/13197 AB - 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 4, and retinoic acid.Materials and Methods: 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, and protamine-1 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-3 and protamine-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.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.Key words: Adipose tissue-derived mesenchymal stem cell, Bone morphogenetic protein 4, Germ-line cells, Retinoic acid, Sertoli cells. ER -