Laboratory of Animal Biotechnology, Federal Rural University of Semiarid, Mossoro, 59625900, Brazil.
J Mol Histol. 2024 Dec;55(6):1199-1209. doi: 10.1007/s10735-024-10259-5. Epub 2024 Sep 9.
Conservation of the genetic diversity through skin and cartilage biobanks represents an essential strategy for maintaining biodiversity. Biobanks for the wild species of the order Rodentia have been little studied. Considering that the cryopreservation technique has specific relationships with the tissue and species of interest, we propose investigating different techniques for preserving tissue integrity and cell viability after cartilage and skin culture from Spix's yellow-toothed cavies. Subsequently, two techniques [solid-surface vitrification (SSV) vs. slow freezing (SF)] were used for cartilage and skin cryopreservation. Tissues not subjected to cryopreservation were used as controls. All tissues were evaluated for morphology and proliferation by histological techniques. Moreover, fragments were cultured, and cells were evaluated for viability, proliferation, metabolism, and apoptosis. Regardless of the cryopreservation technique, no differences were observed for the thickness of the epidermis, dermis, skin, spinous and basal layers, fibroblasts, and proliferative activity regarding the number of nucleolar organizer regions (NOR). SSV ensured better maintenance of epidermal cells, normal chondrocytes, filled gaps, collagen fibers, proliferative activity by NOR area/cell, and reduced perinuclear halos and empty gaps compared to SF. SF ensured the conservation of corneum thickness compared to the control. Although both techniques promoted cell recovery after culture, cells from SF resulted in better subconfluence time and day with cell growth around fragments compared to SSV. In conclusion, both cryopreservation techniques resulted in viable cells after culture. However, SSV promoted better maintenance of tissue morphological integrity, and SF ensured the preservation of all cell quality parameters in Spix's yellow-toothed cavies.
通过皮肤和软骨生物库保护遗传多样性是维持生物多样性的一项重要策略。对啮齿目野生动物生物库的研究甚少。考虑到冷冻保存技术与感兴趣的组织和物种有特定的关系，我们建议研究不同的技术，以保持 Spix 黄牙豚鼠的软骨和皮肤培养后的组织完整性和细胞活力。随后，我们使用了两种技术[固相玻璃化（SSV）与慢速冷冻（SF）]来冷冻保存软骨和皮肤。未进行冷冻保存的组织作为对照。所有组织均通过组织学技术评估形态和增殖情况。此外，还对组织碎片进行了培养，并评估了细胞活力、增殖、代谢和凋亡情况。无论采用哪种冷冻保存技术，表皮、真皮、皮肤、棘突和基底层、成纤维细胞的厚度以及核仁组成区（NOR）的数量与增殖活性均无差异。与 SF 相比，SSV 能更好地维持表皮细胞、正常软骨细胞、填充间隙、胶原纤维、NOR 区域/细胞的增殖活性，并减少核周晕和空化间隙。SF 与对照组相比，能更好地保持角质层厚度。尽管两种技术都能促进细胞在培养后的恢复，但与 SSV 相比，SF 培养的细胞能更快达到亚汇合状态，并且细胞在碎片周围的生长情况也更好。总之，两种冷冻保存技术都能使培养后的细胞保持活力。然而，SSV 能更好地保持组织形态完整性，SF 则能确保 Spix 黄牙豚鼠的所有细胞质量参数得到保存。
