Effect of melatonin on SSCs colony formation in vitro

Spermatogonial stem cells (SSCs) use as an excellent model system for differentiation, development and functioning of testes and provide a source of cells for infertility treatment and production of transgenic animals and recombinant drugs. The aim of present study was to determine the effects of different concentration of melatonin on prepubertal lamb’s spermatogonial colony formation.                                                                                                                                               

 An experimental study was conducted. Each treatment was replicated 5 times. Testicular cells were isolated from testes of prepubertal lambs by two-step enzymatic digestion then cultured for 10 days. Control cells (C) were grown in basic DMEM media with 1% antibiotic and 5% FBS. Treated cells were grown in basic media with 1 µmol/ml (T1) or 1 nmol/ml (T2) melatonin. Culture media was changed every 72h and SSCs colony formation was monitored by inverted microscope. The data were analyzed by one way ANOVA. P value less than 5% was considered statistically signifent.

SSCs were identified by immunocytochemistry assay against PGP9.5.  Number and Surface area of colonies were greater in T1 group than C and T2 groups at day 7 (P=0.04587).

The findings suggest that simultaneous addition of 1 µmol/ml melatonin to culture media may increase In vitro colony formation of prepubertal sheep SSCs. So addition of above dose of melatonin to SSCs culture media are suggested by authors.