title: Effects of testicular interstitial fluid on the proliferation of the mouse spermatogonial stem cells in vitro.
abstract: Spermatogenesis is a process in adult male mammals supported by spermatogonial stem cells (SSCs). The cultivation of SSCs has potential value, for example for the treatment of male infertility or spermatogonial transplantation. Testicular interstitial fluid was added to culture medium to a final concentration of 5, 10, 20, 30 or 40%, in order to investigate its effects on proliferation of mouse SSCs in vitro, Alkaline phosphatase (AKP) assay, reverse transcription polymerase chain reaction (RT-PCR) analysis and indirect immunofluorescence of cells were performed to identify SSCs, and the proliferation rate and diameters of the SSCs colonies were measured. The results showed that the optimal addition of testicular interstitial fluid to culture medium was 30%. When medium supplemented with 30% testicular interstitial fluid was used to culture mouse SSCs, the optimum proliferation rate and diameter of the cell colonies were 72.53% and 249 μm, respectively, after 8 days in culture, values that were significant higher than those found for other groups (P < 0.05). In conclusion, proliferation of mouse SSCs could be promoted significantly by supplementation of the culture medium with 30% testicular interstitial fluid. More research is needed to evaluate and understand the precise physiological role of testicular interstitial fluid during cultivation of SSCs.
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Hua
authors.initials: H
authors.lastname: Shang
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Guang-Xuan
authors.initials: GX
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Jian-Hong
authors.initials: JH
authors.lastname: Hu
authors.affiliation: College of Animal Science and Technology,Northwest A&F University,Yangling,Shaanxi 712100,P.R. China.
authors.forename: Qing-Wang
authors.initials: QW
authors.lastname: Li
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D045604:Extracellular Fluid / Q000502:physiology*; D019084:Fluorescent Antibody Technique, Indirect; D005786:Gene Expression Regulation / Q000187:drug effects; D008297:Male; D008813:Mice, Inbred ICR; D013093:Spermatogonia / Q000166:cytology* / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013737:Testis; D060245:Tetraspanin 29 / Q000378:metabolism
chemical_list: C554870:Cd9 protein, mouse; D003470:Culture Media; D060245:Tetraspanin 29; D000469:Alkaline Phosphatase