Date palm (Phoenix dactylifera L.) pollen (DPP) is widely used as a folk remedy for male infertility treatment, and has well known medicinal effects.
This study aimed to determine the in vitro effects of DPP on the efficiency of neonate mouse spermatogonial stem cells (SSCs) proliferation.
Sertoli and SSCs were isolated from 6-10-days-old mouse testes, and their identity was confirmed using immunocytochemistry against cytokeratin for sertoli cells and PLZF, Oct-4 and CDH-1 for SSCs. Isolated testicular cells were cultured in the absence or presence of 0.06, 0.25 and 0.62 mg/mL concentrations of DPP aqueous extract for 2 weeks. The number and diameter of SSC colonies were assessed during third, 7th, 9th and 14th day of culture, and the expression of the Mvh, GFRα-1 and Oct-4 was evaluated using quantitative PCR at the end of the culture period. The significance of the data was analyzed using ANOVA and paired samples t-test and Tukey and Bonferroni test as post hoc tests at the level of p≤ 0.05.
Pattern assay of colony formation showed that SSCs numbers increased in the present of 0.62 mg/ml concentration of DPP extract with higher slop relative to other groups (P < 0.05). Colony diameters had no significant difference between groups in 3(th), 7(th), 9(th) and 14(th) days after culture. The Mvh and Oct-4 genes expression had no significant difference between groups, while GFRα1 expression was increased significantly in cells treated with 0.06 mg/ml concentration relative to other groups (P < 0.05).
It seems that co-culture of SSCs with sertoli sells in the presence of low doses of DPP can increase SSCs proliferation and keep their stemness state, while higher concentrations can differentiate the treated cells.
Journal of ethnopharmacology. 2016 Apr 12 [Epub ahead of print]
Maryam Mahaldashtian, Majid Naghdi, Mohamad Taghi Ghorbanian, Zohreh Makoolati, Seyedeh Momeneh Mohamadi
Department of Molecular & Cellular Biology, Faculty of Biology, Damghan University, Semnan, Iran; Department of Pharmacology, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran. Electronic address: ., Department of Anatomical Sciences, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran; Department of Pharmacology, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran. Electronic address: ., Department of Molecular & Cellular Biology, Faculty of Biology, Damghan University, Semnan, Iran; Department of Pharmacology, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran. Electronic address: ., Department of Anatomical Sciences, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran; Department of Pharmacology, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran. Electronic address: ., Department of Pharmacology, Faculty of Medicine, Fasa University of Medical Sciences, Fasa,Iran.