title: Improving the Efficacy of Cryopreservation of Spermatogonia Stem Cells by Antioxidant Supplements.
abstract: Cryopreservation of spermatogonial stem cells (SSCs) is an applicable method for young males seeking fertility preservation before starting a treatment. It increases reactive oxygen species (ROS) formation and oxidative stress, which damages cellular structures. In this study, we added two antioxidants, catalase and α-tocopherol (α-TCP), to the basic freezing medium to evaluate their effects on the efficiency of SSCs. SSCs were isolated from testes of 3- to 6-day-old male mice using enzymatic digestion. The enrichment of isolated cells was evaluated by flow cytometry and Stra8 antibody. Catalase (40 μg/mL), or α-TCP (200 μg/mL) was added to the basic freezing medium. The cell viability was evaluated by the methylthiazoltetrazolium (MTT) assay. After thawing, cells were cultured for 1 month, and the expression pattern of specific genes of SSCs and the ability of the cells to restore spermatogenesis were used to determine the efficiency of the cryopreservation method. The survival rate of the frozen cells in the presence of catalase or α-TCP was significantly higher than the control group (p < 0.05). The number of colonies and their diameter measured after 1 month were significantly higher in the antioxidant groups than in the control group (p < 0.05). Gene expression and resumption of spermatogenesis also followed the same pattern. Thus, adding antioxidants to the basic freezing medium can be helpful in increasing the quality and viability of SSCs after cryopreservation. This new approach to stem cells cryopreservation can also be a promising strategy for fertility preservation in patients who suffer from malignancy.
authors.affiliation: 1 Department of Anatomy, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran , 1417613151.
authors.forename: Fereshte
authors.initials: F
authors.lastname: Aliakbari
authors.affiliation: 2 Department of Urology, Shariati Hospital, Tehran University of Medical Sciences , Tehran, Iran , 1417613151.|3 Department of Andrology, Reproductive Biomedicine Research Center, Royan Institute for Stem Cell Biology and Technology , ACECR, Tehran, Iran , 1417613151.
authors.forename: Mohamad Ali Sedighi
authors.initials: MA
authors.lastname: Gilani
authors.affiliation: 1 Department of Anatomy, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran , 1417613151.
authors.forename: Fardin
authors.initials: F
authors.lastname: Amidi
authors.affiliation: 4 Department of Anatomy, School of Medicine, Arak University of Medical Sciences , Arak, Iran , 3133705.
authors.forename: Maryam
authors.initials: M
authors.lastname: Baazm
authors.affiliation: 5 Cellular and Molecular Research Center and Department of Anatomical Sciences, Iran University of Medical Science, Tehran, Iran , 88052965.
authors.forename: Morteza
authors.initials: M
authors.lastname: Korouji
authors.affiliation: 6 Primegen Biotech LLC, Santa Ana, California.
authors.forename: Fariborz
authors.initials: F
authors.lastname: Izadyar
authors.affiliation: 1 Department of Anatomy, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran , 1417613151.
authors.forename: Hosein
authors.initials: H
authors.lastname: Yazdekhasti
authors.affiliation: 1 Department of Anatomy, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran , 1417613151.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Abbasi
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015925:Cryopreservation / Q000379:methods*; D008297:Male; D051379:Mice; D013093:Spermatogonia / Q000166:cytology*; D024502:alpha-Tocopherol / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D024502:alpha-Tocopherol