Biopolymer gels as components of protective medium for cryopreservation of spermatogonial stem cells

Abstract

Biopolymer gels attract a lot of attention in a field of biothechnology due to their excellent compatibility and degradation. Their application is also promising for cryopreservation of spermatogonial stem cells (SSCs) which is so necessary to preserve the fertility of young patients. The aim of the study was to determine the effectiveness of biopolymer gels as a component of cryopreservation medium for SSCs of immature rats at the stage of exposure to cryoprotectants. It was found that 30-min exposure to cryopreservation media based on collagen or fibrin gel with an addition of 5% Me₂SO or 6% glycerol did not lead to significant changes in membrane integrity, cytochrome C content, metabolic, mitochondrial and antioxidant activities in SSCs compared to the control (Leibovitz-based cryomedium). But fibrin gel more than collagen reduced the toxic effects of Me₂SO and glycerol on SSCs increasing exposure time up to 45 min without significant changes in cell viability. The same cryoprotectants in Leibovitz-based media showed significant toxicity starting from the 15th minute of exposure. Necrosis was the main cause of cell death at this stage of cryopreservation in all experimental groups. The obtained results can be used to optimize SSC cryopreservation protocols for further cell autotransplantation for spermatogenesis initiation in boys who undergo gonadotoxic therapy in prepubertal age.

Keywords

Spermatogonial stem cells fibrin gel collagen gel cryopreservation medium exposure apoptosis necrosis

Get full access to this article

View all access options for this article.

References

Haack-Sorensen

Bindslev

Mortensen

, et al. The influence of freezing and storage on the characteristics and functions of human mesenchymal stromal cells isolated for clinical use. Cytotherapy 2007; 9(4): 328–337, DOI: 10.1080/14653240701322235

Sperm banking before gonadotoxic treatment: is it worth the effort?

Asian Journal Andrology 2021; 23(5): 490–494, DOI: 10.4103/aja.aja_16_21

Mahmud

Cryopreservation of Stem Cell Product. In: Chandy

Radhakrishnan

Sukumaran

(eds). Contemporary Bone Marrow Transplantation. Organ and Tissue Transplantation. Cham: Springer, 2020, DOI: 10.1007/978-3-319-64938-2_16-1

Vilaj

Golubić-Ćepulić

Ježek

. Spermatogonial Stem Cell Cryopreservation for Fertility Preservation. In: Virant-Klun

(ed). Stem Cells in Reproductive Tissues and Organs. Stem Cell Biology and Regenerative Medicine. Cham: Humana, 2022, DOI: 10.1007/978-3-030-90111-0_7

Unni

Kasiviswanathan

D’Souza

, et al. Efficient cryopreservation of testicular tissue: effect of age, sample state, and concentration of cryoprotectant. Fertil Steril 2012; 97: 200–208, DOI: 10.1016/j.fertnstert.2011.10.018

Arakawa

Kita

Timasheff

. Protein precipitation and denaturation by dimethyl sulfoxide. Biophys Chem 2007; 131: 62–70, DOI: 10.1016/j.bpc.2007.09.004

Lee

Kim

, et al. Cryopreservation of mouse spermatogonial stem cells in dimethylsulfoxide and polyethylene glycol. Biol Reprod 2013; 89: 109, DOI: 10.1095/biolreprod.113.111195

Kim

Lee

Kim

, et al. Cryopreservation of putative pre-pubertal bovine spermatogonial stem cells by slow freezing. Cryobiology 2015; 70(2): 175–183, DOI: 10.1016/j.cryobiol.2015.02.007

Izadyar

Matthijs‐Rijsenbilt

Den Ouden

, et al. Development of a cryopreservation protocol for type A spermatogonia. J Andrology 2002; 23(4): 537–545, DOI: 10.1002/j.1939-4640.2002.tb02276.x

, et al. Insights into the crystallization and vitrification of cryopreserved cells. Cryobiology 2022; 106: 13–23, DOI: 10.1016/j.cryobiol.2022.04.008

. Review paper: critical issues in tissue engineering: biomaterials, cell sources, angiogenesis, and drug delivery systems. J Biomaterials Applications 2011; 26(4): 383–417, DOI: 10.1177/0885328211408946

, et al. Hydrogels based on collagen and fibrin – frontiers and applications. BioNanoMaterials 2016; 17(1–2). DOI: 10.1515/bnm-2015-0025

13.

Spicer

. Hydrogel scaffolds for tissue engineering: The importance of polymer choice. Polym Chem 2019; 11(2): 184–219, DOI: 10.1039/c9py01021a

, et al. A tough act to follow: collagen hydrogel modifications to improve mechanical and growth factor loading capabilities. Mater Today. Bio 2021; 10: 100098, DOI: 10.1016/j.mtbio.2021.100098

, et al. Potential of Fibrin Glue and Mesenchymal Stem Cells (MSCs) to Regenerate Nerve Injuries: A Systematic Review. Cells 2022; 11(2): 221, DOI: 10.3390/cells11020221

16.

Lee

, et al. Enhanced cellular cryopreservation by biopolymer-associated suppression of RhoA/ROCK Signaling Pathway. Materials 2021; 14(20): 6056, DOI: 10.3390/ma14206056

. Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review. Biomacromolecules 2011; 12(5): 1387–1408, DOI: 10.1021/bm200083n

, et al. Cryopreservation: An Overview of Principles and Cell-Specific Considerations. Cel Transplantation 2021; 30: 963689721999617, DOI: 10.1177/0963689721999617

, et al. Cryopreservation of Tissue-Engineered Scaffold-Based Constructs: from Concept to Reality. Advance online publication, 2021. DOI: 10.1007/s12015-021-10299-4

, et al. Cryopreserved fragments of testicular seminiferous tubules of rats as a source of spermatogonial stem cells. Cell Organ Transpl 2021; 9(1): 36–42, , DOI: 10.22494/cot.v9i1.120

21.

Best

. Cryoprotectant Toxicity: Facts, Issues, and Questions. Rejuvenation Research 2015; 18(5): 422–436, DOI: 10.1089/rej.2014.1656

22.

Gao

Tian

, et al. Advanced biomaterials in cell preservation: Hypothermic preservation and cryopreservation. Acta Biomater 2021; 131: 97–116, DOI: 10.1016/j.actbio.2021.07.001

23.

Stacey

Masters

. Cryopreservation and banking of mammalian cell lines. Nat Protoc 2008; 3(12): 1981–1989, DOI: 10.1038/nprot.2008.190

, et al. Mitochondrial Membrane Potential Predicts 4-Hour Sperm Motility. Biomedicines 2020; 8(7): 196, DOI: 10.3390/biomedicines8070196

, et al. Association of active caspase 8 with the mitochondrial membrane during apoptosis: potential roles in cleaving BAP31 and caspase 3 and mediating mitochondrion-endoplasmic reticulum cross talk in etoposide-induced cell death. Mol Cellular Biology 2004; 24(15): 6592–6607, DOI: 10.1128/MCB.24.15.6592-6607.2004

. Analysis of the Mitochondrial Membrane Potential Using the Cationic JC-1 Dye as a Sensitive Fluorescent Probe. Bio-protocol 2019; 9(1): e3128, DOI: 10.21769/BioProtoc.3128

, et al. Influence of a Virgin Olive Oil versus Butter Plus Cholesterol-Enriched Diet on Testicular Enzymatic Activities in Adult Male Rats. Int Journal Molecular Sciences 2017; 18(8): 1701, DOI: 10.3390/ijms18081701

. The Pentose Phosphate Pathway in Yeasts-More Than a Poor Cousin of Glycolysis. Biomolecules 2021; 11(5): 725, DOI: 10.3390/biom11050725

, et al. G6PD overexpression protects from oxidative stress and age-related hearing loss. Aging Cell 2020; 19(12): e13275, DOI: 10.1111/acel.13275

, et al. Glucose-6-phosphate dehydrogenase inhibition attenuates acute lung injury through reduction in NADPH oxidase-derived reactive oxygen species. Clin Experimental Immunology 2018; 191(3): 279–287, DOI: 10.1111/cei.13097

, et al. Cryopreservation of Rat Seminiferous Tubules Using Biopolymers and Slow Non-Controlled Rate Cooling. Probl Cryobiology Cryomedicine 2019; 28(4): 278–292, DOI: 10.15407/cryo28.04.278

, et al. Cryopreservation of Human Ovarian Tissue. Cell Tissue Banking 2006; 7: 123–133, DOI: 10.1007/s10561-005-1968-8

, et al. Exposure of Seminiferous Tubules of Immature Rats to Cryoprotective Media of Various Compositions. Probl Cryobiology Cryomedicine 2017; 27(3): 203–218, DOI: 10.15407/cryo27.03.203

, et al. Trehalose to cryopreserve human pluripotent stem cells. Stem Cell Research 2018; 31: 102–112, DOI: 10.1016/j.scr.2018.07.021

, et al. Hydrogel Microfiber Encapsulation Enhances Cryopreservation of Human Red Blood Cells with Low Concentrations of Glycerol. Biopreservation and Biobanking 2020; 18(3): 228–234, DOI: 10.1089/bio.2020.0003

, et al. Gelatin carrageenan sericin hydrogel composites improves cell viability of cryopreserved SaOS-2 cells. Int Journal Biological Macromolecules 2020; 154: 606–620, DOI: 10.1016/j.ijbiomac.2020.03.039

. Biopolymer-based growth factor delivery for tissue repair: from natural concepts to engineered systems. Tissue Eng B: Rev 2009; 15(3): 263–289, DOI: 10.1089/ten.teb.2008.0668

. Advancing biomaterials of human origin for tissue engineering. Prog Polymer Science 2016; 53: 86–168, DOI: 10.1016/j.progpolymsci.2015.02.004

. Electrospraying of polymers with therapeutic molecules: State of the art. Prog Polymer Science 2012; 3711: 1510–1551, DOI: 10.1016/j.progpolymsci.2012.03.002

, et al. Hydrogel Cryopreservation System: An Effective Method for Cell Storage. Int Journal Molecular Sciences 2018; 19(11): 3330, DOI: 10.3390/ijms19113330

, et al. Optimizing Blended Collagen-Fibrin Hydrogels for Cardiac Tissue Engineering with Human iPSC-derived Cardiomyocytes. ACS Biomater Sci Eng 2019; 5(2): 887–899, https://doi.org/10.1021/acsbiomaterials.8b01112

42.

Coffin

Gaudette

. Aprotinin extends mechanical integrity time of cell‐seeded fibrin sutures. J Biomed Mater Res A 2016; 104(9): 2271–2279, https://doi.org/10.1002/jbm.a.35754

43.

Zeng

Yin

Zhang

, et al. A Supramolecular Gel Approach to Minimize the Neural Cell Damage during Cryopreservation Process. Macromolecular Biosci 2015; 16(3): 363–370, https://doi:10.1002/mabi.201500277

44.

Thuwanut

Comizzoli

Pimpin

, et al. Influence of hydrogel encapsulation during cryopreservation of ovarian tissues and impact of post-thawing in vitro culture systems in a research animal model. Clin Exp Reprod Med 2021; 48(2): 111–123, https://doi.org/10.5653/cerm.2020.04056

45.

Bock

Tait

Mitochondria as multifaceted regulators of cell death. Nat Reviews. Mol Cell Biology 2020; 21(2): 85–100, DOI: 10.1038/s41580-019-0173-8

, et al. Cytochrome C speeds up caspase cascade activation by blocking 14-3-3ε-dependent Apaf-1 inhibition. Cel Death Disease 2018; 9(3): 1–12, DOI: 10.1038/s41419-018-0408-1

47.

Trubiani

Pieri