Cryopreservation of Rat Seminiferous Tubules Using Biopolymers and Slow Non-Controlled Rate Cooling
Keywords:seminiferous tubules, immature rats, cryoprotectors, bovine serum albumin, collagen gel, ï¬brin gel
One of the current trends in biomedical research is the use of novel biological products and cryotechniques to maintain fertility, especially in case of planned cytotoxic therapy. Existing protocols of testicular tissue cryopreservation require further optimization and improvement. This study represents a comparative evaluation of the effect of biopolymers (bovine serum albumin, collagen and fibrin gel) in combination with either 0.6M dimethylsulfoxide or 0.7 M glycerol on morphofunctional characteristics of seminiferous tubules of immature rats after cryopreservation using slow non-controlled cooling down to 70Â°C for 40 min with following immersing the samples into liquid nitrogen. Based on the results of the analysis of viability, total activity of lactate dehydrogenase, MTT-test and histomorphometric data (cell retraction degree, epithelium desquamation frequency, number of cells with condensed nuclues, existingÂ spermatogenic layer deffects), the combination of the fibrin gel with a 0.7 M glycerol has been found to be the most optimal among all the studied combinations of biopolymers and cryoprotectants. The obtained data can be used for substantiation and development of effective cryopreservation methods for human seminiferous tubules using biopolymers.
Probl Cryobiol Cryomed 2018; 28(4): 278-292
Allenspach AL, Kraemer TG. Ice crystal patterns in artiï¬cial gels of extracellular matrix molecules after quick-freezing and freeze-substitution. Cryobiology. 1989; 26(2): 170â€“9.
Belous AM, Gricshenko VI. [Cryobiology]. Kiev: Naukova dumka, 1994. 428 p. Russian.
Bystrova OV, Kalugina AS, Tsybatova EV, et al. [Methods for recovering fertility in cancer patients]. Prakt Onkologiya. 2009; 10(4): 245â€“53. Russian.
Campion SN, Carvallo FR, Chapin RE, et al. Comparative assessment of the timing of sexual maturation in male Wistar Han and Sprague-Dawley rats. Reprod Toxicol. 2013; 38(7):16â€“24.
Chandrakasan G, Torchia DA, Piez KA. Preparation of intact monomeric collagen from rat tail tendon and skin and the structure of the nonhelical ends in solution. J Biol Chem. 1976;251: 6062â€“7. PubMed
Chiti MC, Dolmans MM, Donnez J, Amorim CA. Fibrin in reproductive tissue engineering: a review on its application as a biomaterial for fertility preservation. Annals of biomedical engineering. 2017; 45(7): 1650â€“63.
Dimasi L. Meeting increased demands on cell-based processes by using deï¬ned media supplements. BioProcess International. 2011; 9(8): 48â€“58.
European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientiï¬c Purposes. European Treaty Series No. 123 Strasbourg, 18.III.1986. Strasbourg: Council of Europe; 1986. 53p. [Cited 01.02.2018] Available from: https://rm.coe.int/CoERMPublicCommonSearchServices/ DisplayDCTMContent?documentId=090000168007a67b.
Easley C, Simerly C, Schatten G. Stem cell therapeutic pos- sibilities: future therapeutic options for male-factor and female-factor infertility? Reprod Biomed Online [Internet]. 2013; 27(1): 75â€“80. [Cited 01.02.2018] Available from: https://www.sciencedirect.com/science/article/pii/ S1472648313001296?via%3Dihub. CrossRef
Farrant J. Is there a common mechanism of protection of living cells by polyvinylpyrrolidone and glycerol during freezing? Nature. 1969; 222: 1175â€“6. CrossRef
Ginsburg ES, Yanushpolsky EH, Jackson KV. In vitro fertilization for cancer patients and survivors. Fertility and Sterility. 2001; 75(4): 705â€“10.
Grill G, Porcellini A, Lucarelli G. Role of serum on cryopreservation and subsequent viability of mouse bone marrow hemopoetic stem cells. Cryobiology. 1980; 17(5): 516â€“20.
Jahnukainen K, Ehmcke J, SÐ¾der O, et al. Clinical potential and putative risks of fertility preservation in children utilizing gonadal tissue or germline stem cells. Pediatr Res. 2006; 59: 40Râ€“47R. CrossRef
Kaidasheva IP. [Methods of clinical and experimental research in medicine]. Poltava: Polimet; 2003. 320 p. Ukrainian.
Keros V, Rosenlund B, Hultenby K, et al. Optimizing cryopre- servation of human testicular tissue: comparison of protocols with glycerol, propanediol and dimethylsulphoxide as cryoprotectants. Hum Reprod. 2005; 20(6): 1676â€“87.
Koebe HG, Dunn JC, Toner M, et al. A new approach to the cryopreservation of hepatocytes in a sandwich culture conï¬guration. Cryobiology. 1990; 27(5): 576â€“84.
Kvist K, Thorup J, Byslov AG, et al. Cryopreservation of intact testicular tissue from boys with cryptorchidism. Hum Reprod. 2006; 21(2): 484â€“91.
Milazzo JP, Travers A, Bironneau A, et al. Rapid screening of cryopreservation protocols for murine prepubertal testicular tissue by histology and PCNA immunostaining. J Androl. 2010; 31(6): 617â€“30.
Miyamoto Y, Enosawa S, Takeuchi T, et al. Cryopreservation in situ of cell monolayers on collagen vitrigel membrane culture substrata: ready-to-use preparation of primary hepatocytes and ES cells. Cell Transplant. 2009; 18(5): 619â€“26.
Mossman T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2): 55â€“63. CrossRef
Poels J, Abou-Ghannam G, Herman S, et al. In search of better spermatogonial preservation by supplementation of cryopreserved human immature testicular tissue xenografts with n-acetylcysteine and testosterone. Front Surg [Internet]. 2014; 1: 47. [Cited 01.02.2018] Available from: https://www. frontiersin.org/articles/10.3389/fsurg.2014.00047/full. CrossRef
Takahashi T, Hirsh A, Erbe E, Williams RJ. Mechanism of cryoprotection by extracellular polymeric solutes. Biophys J. 1988; 54(3): 509â€“18.
Volkova NA, Pavlovich EV, Gapon AA, Nikolov OT. Eï¬€ects of millimeter-wave electromagnetic exposure on the morphology and function of human cryopreserved spermatozoa. Bull Exp Biol Med. 2014; 157(5): 574â€“6.
Volkova NO, Yukhta MS, Chernyshenko LG, et al. Exposure of seminiferous tubules of immature rats to cryoprotective media of various compositions. Probl Cryobiol Cryomed. 2017; 27(3): 203â€“18.
Volkova NÐ, Yukhta MS, Goltsev A. Mesenchymal stem cells in restoration of fertility at experimental pelvic inï¬‚ammatory disease. Stem cells international [Internet]. 2017; Article ID 2014132. [Cited 01.02.2018] Available from: https://www. hindawi.com/journals/sci/2017/2014132/. CrossRef
Williams RJ. The surface activity of PVP and other polymers and their antihemolytic capacity. Cryobiology. 1983; 20: 521â€“6.
Wyns C, Curaba M, Vanabelle B, et al. Options for fertility preservation in prepubertal boys. Hum Reprod Update. 2010;16(3): 312â€“28.
Yudintseva N, Pleskach N, Smagina L, et al. Reconstruction of connective tissue from ï¬brin-based dermal equivalent transplanted to animals with experimental wounds. Cell and Tissue Biology. 2010; 4(5): 476â€“80.
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