Introduction: Mouse spermatogonial stem cells (SSCs) can be cryopreserved for long periods while preserving their spermatogenic ability. Although cryopreservation has been found to increase reactive oxygen species (ROS) formation that damages cellular structures. In the present study, we added vitamin E to the basic freezing medium in order to evaluate its effect on the efficiency of spermatogonial stem cells.
Methods: SSCs isolated from testes of 6 days old male mice by enzymatic digestion. Vitamin E 100, 200, 400 µg/mL was added to the basic freezing medium. The cell viability was evaluated by MTT assay. After thawing, SSCs were cultured for 1 month and the expression pattern of specific genes of SSCs measured by real-time PCR technique.
Results: The survival rate of the freeze cells in the presence of vitamin E was significantly higher than the control group (p<0.05). The number of colonies and their diameter measured after one month were significantly higher in the vitamin E groups than in the control group (p≤0.05).
Conclusion: Adding vitamin E to the basic freezing medium thus can be helpful in increasing the quality and viability of SSCs after cryopreservation.
Sato T, Sakuma T, Yokonishi T, Katagiri K, Kamimura S, Ogonuki N, et al. Genome editing in mouse spermatogonial stem cell lines using TALEN and double-nicking CRISPR/Cas9. Stem Cell Reports. 2015;5(1):75-82.
Vansandt LM, Livesay JL, Dickson MJ, Li L, Pukazhenthi BS, Keefer CL. Conservation of spermatogonial stem cell marker expression in undifferentiated felid spermatogonia. Theriogenology. 2016.
Kanatsu-Shinohara M, Morimoto H, Shinohara T. Enrichment of mouse spermatogonial stem cells by melanoma cell adhesion molecule expression. Biology of reproduction. 2012;87(6):139.
Takashima S, Kanatsu-Shinohara M, Tanaka T, Morimoto H, Inoue K, Ogonuki N, et al. Functional differences between GDNF-dependent and FGF2-dependent mouse spermatogonial stem cell self-renewal. Stem cell reports. 2015;4(3):489-502.
Wyns C, Curaba M, Petit S, Vanabelle B, Laurent P, Wese J-F, et al. Management of fertility preservation in prepubertal patients: 5 years’ experience at the Catholic University of Louvain. Human reproduction. 2011:deq387.
Heidari B, Rahmati-Ahmadabadi M, Akhondi MM, Zarnani AH, Jeddi-Tehrani M, Shirazi A, et al. Isolation, identification, and culture of goat spermatogonial stem cells using c-kit and PGP9. 5 markers. Journal of assisted reproduction and genetics. 2012;29(10):1029-38.
Goossens E, Van Saen D, Tournaye H. Spermatogonial stem cell preservation and transplantation: from research to clinic. Human Reproduction. 2013;28(4):897-907.
Avarbock MR, Brinster CJ, Brinster RL. Reconstitution of spermatogenesis from frozen spermatogonial stem cells. Nature medicine. 1996;2(6):693.
Lushchak VI. Environmentally induced oxidative stress in aquatic animals. Aquatic Toxicology. 2011;101(1):13-30.
Elgaml SA, Khalil R, Hashish EA, El-Murr A. Protective Effects of Selenium and Alpha-Tocopherol against Lead-Induced Hepatic and Renal Toxicity in Oreochromis Niloticus. Journal of Aquaculture Research & Development. 2015;2015.
Patel S, Hans MK, Chander S, Ahluwalia AS. Antioxidants in Endodontics: A Strategic Review. Journal of clinical and diagnostic research: JCDR. 2015;9(5):ZE12.
Abadie‐Guedes R, Bezerra RdS, Guedes RCA. Alpha‐Tocopherol Counteracts the Effect of Ethanol on Cortical Spreading Depression in Rats of Various Ages, With and Without Ethanol Abstinence. Alcoholism: Clinical and Experimental Research. 2016.
Ibrahim M, Hassan W, Meinerz DF, de Oliveira Leite G, Nogueira CW, Rocha JB. Ethanol-induced oxidative stress: The role of binaphthyl diselenide as a potent antioxidant. Biological trace element research. 2012;147(1-3):309-14.
Aliakbari F, Yazdekhasti H, Abbasi M, Hajian Monfared M, Baazm M. Advances in cryopreservation of spermatogonial stem cells and restoration of male fertility. Microscopy research and technique. 2016;79(2):122-9.
Altman E, Yango P, Moustafa R, Smith JF, Klatsky PC, Tran ND. Characterization of human spermatogonial stem cell markers in fetal, pediatric, and adult testicular tissues. Reproduction. 2014;148(4):417-27.
Kanatsu-Shinohara M, Morimoto H, Shinohara T. Fertility of Male Germline Stem Cells Following Spermatogonial Transplantation in Infertile Mouse Models. Biology of Reproduction. 2016:biolreprod. 115.137869.
Izadyar F, Matthijs‐Rijsenbilt J, OUDEN K, CREEMERS LB, WOELDERS H, ROOIJ DG. Development of a cryopreservation protocol for type A spermatogonia. Journal of andrology. 2002;23(4):537-45.
Rhee SG, Chang T-S, Jeong W, Kang D. Methods for detection and measurement of hydrogen peroxide inside and outside of cells. Molecules and cells. 2010;29(6):539-49.
Koruji M, Movahedin M, Mowla SJ, Gourabi H. Colony formation ability of frozen thawed spermatogonial stem cell from adult mouse. Iranian Journal of Reproductive Medicine. 2007;5(3):109-15.
Shinohara T, Inoue K, Ogonuki N, Kanatsu-Shinohara M, Miki H, Nakata K, et al. Birth of offspring following transplantation of cryopreserved immature testicular pieces and in-vitro microinsemination. Human Reproduction. 2002;17(12):3039-45.
Jahnukainen K, Ehmcke J, Hergenrother SD, Schlatt S. Effect of cold storage and cryopreservation of immature non-human primate testicular tissue on spermatogonial stem cell potential in xenografts. Human Reproduction. 2006;22(4):1060-7.
Hermann BP, Sukhwani M, Lin CC, Sheng Y, Tomko J, Rodriguez M, et al. Characterization, cryopreservation, and ablation of spermatogonial stem cells in adult rhesus macaques. Stem cells. 2007;25(9):2330-8.
Valentino S, Ghelfi M, Zunica E, Stamper M, Hickman S, Hwang S, et al. Antioxidant independent actions of vitamin E in modulating gene expression. Free Radical Biology and Medicine. 2018;128:S58-S9.
Zarandi NP, Galdon G, Kogan S, Atala A, Sadri-Ardekani H. Cryostorage of immature and mature human testis tissue to preserve spermatogonial stem cells (SSCs): a systematic review of current experiences toward clinical applications. Stem cells and cloning: advances and applications. 2018;11:23.
Oatley J. Cryopreserving and thawing spermatogonial stem cells. Cold Spring Harbor Protocols. 2017;2017(4):pdb. prot094219.
Um H-D. Bcl-2 family proteins as regulators of cancer cell invasion and metastasis: a review focusing on mitochondrial respiration and reactive oxygen species. Oncotarget. 2016;7(5):5193.