Gastal G D A, Alves B G, Alves K A, Souza M E M, Vieira A D, Varela A S, Figueiredo J R, Feugang J M, Lucia T, Gastal E L
Department of Animal ScienceFood and Nutrition, Southern Illinois University, Carbondale, Illinois, USA.
Laboratory of Animal ReproductionFaculty of Veterinary Medicine.
Reproduction. 2017 May;153(5):577-587. doi: 10.1530/REP-16-0621. Epub 2017 Feb 28.
The method of transportation and the conditions imposed on the ovarian tissue are pivotal aspects for the success of ovarian tissue cryopreservation (OTC). The aim of this study was to evaluate the effect of the size of the ovarian tissue (e.g. whole ovary, biopsy size and transplant size) during different times of storage (0, 6, 12 and 24 h) on the structural integrity of equine ovarian tissue transported at 4°C. Eighteen pairs of ovaries from young mares (<10 years old) were harvested in a slaughterhouse and processed to simulate the fragment sizes (biopsy and transplant size groups) or kept intact (whole ovary group) and stored at 4°C for up to 24 h in α-MEM-enriched solution. The effect of the size of the ovarian tissue was observed on the morphology of preantral follicles, stromal cell density, DNA fragmentation and mitochondrial membrane potential. The results showed that (i) biopsy size fragments had more morphologically normal preantral follicles after 24 h of storage at 4°C; (ii) mitochondrial membrane potential was the lowest during each storage time when the whole ovary was used; (iii) DNA fragmentation rate in the ovarian cells of all sizes of fragments increased as storage was prolonged and (iv) transplant size fragments had increased stromal cell density during storage at cool temperature. In conclusion, the biopsy size fragment was the best to preserve follicle morphology for long storage (24 h); however, transportation/storage should be prior determined according to the distance (time of transportation) between patient and reproduction centers/clinics.
卵巢组织的运输方式和所施加的条件是卵巢组织冷冻保存（OTC）成功的关键因素。本研究的目的是评估在4°C运输时，不同储存时间（0、6、12和24小时）下卵巢组织大小（如整个卵巢、活检大小和移植大小）对马卵巢组织结构完整性的影响。从屠宰场采集18对年轻母马（<10岁）的卵巢，进行处理以模拟不同的碎片大小（活检和移植大小组）或保持完整（整个卵巢组），并在富含α-MEM的溶液中于4°C储存长达24小时。观察卵巢组织大小对腔前卵泡形态、基质细胞密度、DNA片段化和线粒体膜电位的影响。结果表明：（i）在4°C储存24小时后，活检大小的碎片中有更多形态正常的腔前卵泡；（ii）使用整个卵巢时，每个储存时间的线粒体膜电位最低；（iii）随着储存时间延长，所有大小碎片卵巢细胞中的DNA片段化率均增加；（iv）在低温储存期间，移植大小的碎片基质细胞密度增加。总之，活检大小的碎片最适合长时间（24小时）保存卵泡形态；然而，应根据患者与生殖中心/诊所之间的距离（运输时间）预先确定运输/储存方式。
