Bono-Mestre C, Cardona-Costa J, García-Ximénez F
Laboratory of Animal Reproduction and Biotechnology (LARB-UPV), Polytechnic University of Valencia, Valencia, Spain.
Cryo Letters. 2009 Mar-Apr;30(2):148-52.
In this work, three cryopreservation procedures were tested in order to obtain efficiently viable testicular cells after cryopreservation. Testicular cells of Wild type zebrafish males were frozen using an equilibrium protocol and testicular tissue fragments were cryopreserved with equilibrium freezing and vitrification procedures. Results showed that vitrification was significantly more efficient than freezing in terms of final cell survival (cell freezing: 14.4%, tissue freezing: 77.4%-85.5%, tissue vitrification: 94%). It must be noted that, in live cells, the presence of pseudopodia was frequently observed, which indicated their spermatogonial nature. Based on these results, the authors suggest that vitrification, with the subsequent elimination of connective tissue after warming, offers the best combination to rescue live testicular cells as a genetic conservation procedure in zebrafish.
在这项工作中,测试了三种冷冻保存程序,以便在冷冻保存后高效获得有活力的睾丸细胞。使用平衡方案冷冻野生型斑马鱼雄性的睾丸细胞,并采用平衡冷冻和玻璃化程序冷冻保存睾丸组织碎片。结果表明,就最终细胞存活率而言,玻璃化显著比冷冻更有效(细胞冷冻:14.4%,组织冷冻:77.4%-85.5%,组织玻璃化:94%)。必须指出的是,在活细胞中,经常观察到伪足的存在,这表明它们具有精原细胞的性质。基于这些结果,作者建议,玻璃化结合复温后结缔组织的后续清除,为拯救活的睾丸细胞作为斑马鱼的一种基因保存程序提供了最佳组合。