title: Does prepubertal testicular tissue vitrification influence spermatogonial stem cells (SSCs) viability?
abstract: OBJECTIVE
  Testicular cryopreservation prior to chemotherapy or radiotherapy in children with cancer is one of the ways to preserve fertility. However, cryopreservation may cause damage to the testicular parenchyma cells. The objective of this study was to investigate effects of vitrification on the intracellular LDH leakage, cell cycle/apoptotic responses and apoptosis-related gene expression patterns in the spermatogonial stem cells (SSCs) obtained from the vitrified testis.
  
  
  METHODS
  The testes of the mice pups (6-day-old, BALB/c) both vitrified and fresh groups were digested with enzymes (collagenase, DNaseΙ, trypsin-EDTA) to disperse the cells. The SSCs, type A, were isolated from the rest of testicular cells by MACS. The amount of damage to the SSCs immediately was evaluated by Cytotoxicity assay, Flow cytometry assay and Real-time PCR.
  
  
  RESULTS
  The intracellular LDH leakage in the SSCs,harvested from the vitrified testes, was less reported compared with the fresh ones. Moreover, the percentage of apoptotic and necrotic SSCs obtained from the vitrified testes was lower than that of yielded from the fresh samples. Also, the apoptosis-related genes of the SSCs,collected from the vitrified testes, changed their expression profile as increasing P53 and BCL-2 expression levels and decreasing Bax and Fas expression levels.
  
  
  CONCLUSIONS
  The study indicates that vitrification of prepubertal testicular tissue does not increase the expression profile of apoptosis-related genes such as Bax and Fas in the testicular SSCs consistent with diminished cell apoptotic/necrotic responses and no increasing intracellular LDH leakage.
authors.affiliation: Department of Anatomy, Lorestan University of Medical Sciences, Khorramabad, Iran.
authors.forename: Mohammadreza
authors.initials: M
authors.lastname: Gholami
authors.forename: Masoud
authors.initials: M
authors.lastname: Hemadi
authors.forename: Ghasem
authors.initials: G
authors.lastname: Saki
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Zendedel
authors.forename: Ali
authors.initials: A
authors.lastname: Khodadadi
authors.forename: Javad
authors.initials: J
authors.lastname: Mohammadi-Asl
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002453:Cell Cycle; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D053595:Embryonic Stem Cells / Q000378:metabolism*; D059247:Fertility Preservation / Q000379:methods*; D020869:Gene Expression Profiling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D013093:Spermatogonia / Q000166:cytology*; D013737:Testis; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis; D058989:Vitrification; D051028:bcl-2-Associated X Protein / Q000096:biosynthesis; D019014:fas Receptor / Q000096:biosynthesis
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D019014:fas Receptor; D007770:L-Lactate Dehydrogenase