Division of Reproductive Genetics, Department of Reproductive Science, Kasturba Medical College, Manipal, Manipal Academy of Higher Education, Manipal, 576104, Karnataka, India.
Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences, Manipal Academy of Higher Education, Manipal, 576104, Karnataka, India.
Reprod Sci. 2024 Nov;31(11):3532-3548. doi: 10.1007/s43032-024-01688-4. Epub 2024 Sep 19.
Cryopreservation of testicular tissue holds an important role in the field of fertility preservation, particularly for prepubertal boys diagnosed with cancer. However, prepubertal testicular tissue cryopreservation is still considered to be in the experimental stage necessitating the refinement of cryopreservation protocol. Considering the fact that loss of membrane lipids is the primary cause of freeze-thaw-induced loss of testicular cell functions, in this study, we explored the beneficial properties of exogenous supplementation of membrane lipids in the form of liposomes in enhancing the cryosurvival of prepubertal testicular tissue. The freezing medium supplemented with liposomes (prepared from soy lecithin, phosphatidylethanolamine, phosphatidylserine, and cholesterol) was used for the experiments. Prepubertal testicular tissues from Swiss albino mice were cryopreserved in a liposome-containing freezing medium (LFM) composed of 0.25 mg/mL liposomes, 5% DMSO, and 30% FCS in the DMEM/F12 medium using a slow freezing protocol. The tissues were thawed and assessed for various testicular cell functions. Freezing in LFM mitigated the loss of viability, decreased malondialdehyde level (p < 0.05), and reduced apoptosis (p < 0.05) in the testicular cells compared to the testicular tissue cryopreserved in the control freezing medium (CFM). Further, DMSO (5%) appears to be the ideal penetrating cryoprotectant for prepubertal testicular tissue cryopreservation with liposome-based freezing medium. Similar enhancement in cryosurvival of cells was observed in adult human testicular tissue frozen with LFM. These findings highlight the translational value of liposome-based freezing medium in the cryopreservation of testicular tissue of prepubertal boys undergoing chemotherapy.
冷冻保存睾丸组织在生育力保存领域具有重要作用，特别是对于被诊断患有癌症的青春期前男孩。然而，青春期前睾丸组织冷冻保存仍被认为处于实验阶段，需要完善冷冻保存方案。鉴于膜脂损失是冻融导致睾丸细胞功能丧失的主要原因，在本研究中，我们探讨了外源性补充脂质体形式的膜脂在增强青春期前睾丸组织冷冻保存中的有益特性。冷冻培养基中添加了脂质体（由大豆卵磷脂、磷脂酰乙醇胺、磷脂酰丝氨酸和胆固醇制成），用于实验。将瑞士白化病小鼠的青春期前睾丸组织在含有脂质体的冷冻培养基（LFM）中冷冻保存，该培养基由 0.25mg/mL 脂质体、5%DMSO 和 30%FCS 在 DMEM/F12 培养基中组成，采用缓慢冷冻方案。将组织解冻并评估各种睾丸细胞功能。与在对照冷冻培养基（CFM）中冷冻保存的睾丸组织相比，LFM 中的冷冻可减轻睾丸细胞活力的丧失，降低丙二醛水平（p<0.05）并减少细胞凋亡（p<0.05）。此外，对于青春期前睾丸组织的冷冻保存，5%DMSO 似乎是理想的渗透冷冻保护剂，与基于脂质体的冷冻培养基相结合。在使用 LFM 冷冻的成人人类睾丸组织中也观察到了类似的细胞冷冻保存效果增强。这些发现突出了基于脂质体的冷冻培养基在化疗中青春期前男孩睾丸组织冷冻保存中的转化价值。
