title: In vitro effects of date palm (Phoenix dactylifera L.) pollen on colonization of neonate mouse spermatogonial stem cells.
abstract: BACKGROUND
  Date palm (Phoenix dactylifera L.) pollen (DPP) is widely used as a folk remedy for male infertility treatment, and has well known medicinal effects.
  
  
  OBJECTIVE
  This study aimed to determine the in vitro effects of DPP on the efficiency of neonate mouse spermatogonial stem cells (SSCs) proliferation.
  
  
  METHODS
  Sertoli and SSCs were isolated from 6 to 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 0.62mg/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.
  
  
  RESULTS
  Pattern assay of colony formation showed that SSCs numbers increased in the present of 0.62mg/ml concentration of DPP extract with higher slop relative to other groups (P <0.05). Colony diameters had no significant difference between groups in 3th, 7th, 9th and 14th 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 0.06mg/ml concentration relative to other groups (P<0.05).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Molecular & Cellular Biology, Faculty of Biology, Damghan University, Semnan, Iran. Electronic address: mahaldashtian.m@gmail.com.
authors.forename: Maryam
authors.initials: M
authors.lastname: Mahaldashtian
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Fasa University of Medical Sciences, Fasa, Iran. Electronic address: majidnaghdi@yahoo.com.
authors.forename: Majid
authors.initials: M
authors.lastname: Naghdi
authors.affiliation: Department of Molecular & Cellular Biology, Faculty of Biology, Damghan University, Semnan, Iran. Electronic address: mt.ghorbanian000@gmail.com.
authors.forename: Mohamad Taghi
authors.initials: MT
authors.lastname: Ghorbanian
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Fasa University of Medical Sciences, Fasa, Iran. Electronic address: zohreh1438@yahoo.com.
authors.forename: Zohreh
authors.initials: Z
authors.lastname: Makoolati
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mansoureh
authors.initials: M
authors.lastname: Movahedin
authors.affiliation: Department of Anatomical Sciences, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran.
authors.forename: Seyedeh Momeneh
authors.initials: SM
authors.lastname: Mohamadi
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D064200:Cdh1 Proteins / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D053487:DEAD-box RNA Helicases / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000235:genetics; D008297:Male; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D065749:Phoeniceae; D010936:Plant Extracts / Q000494:pharmacology*; D011058:Pollen / Q000737:chemistry*; D012708:Sertoli Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013093:Spermatogonia / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism
chemical_list: D064200:Cdh1 Proteins; C577585:Fzr1 protein, mouse; C494988:Gfra1 protein, mouse; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D050814:Octamer Transcription Factor-3; D010936:Plant Extracts; C494174:Pou5f1 protein, mouse; C090864:Ddx4 protein, mouse; D053487:DEAD-box RNA Helicases
keywords: Co-culture; Collagenase (PubChem CID: 18680304) Hyaloronidase (PubChem CID: 91820602) Triton X-100 (PubChem CID: 5590); Colonization; Date palm pollen; Sertoli cell; Spermatogonial stem cell