Long-Time Cooling before Cryopreservation Decreased Translocation of Phosphatidylserine (Ptd-L-Ser) in Human Ovarian Tissue

摘要: Objectives To translocation (externalization) of phosphatidylserine lead at least the five negative effects observed during cells cryopreservation: hypoxia, increasing intracellular Ca2+, osmotic disruption cellular membranes, generation reactive oxygen species (ROS) and lipid peroxidation. The aim this study was to test intensiveness immediately after thawing 45 d xenografting human ovarian tissue, which either frozen just operative removal from patient or cooled before cryopreservation 5°C for 24 h then frozen. Materials Methods Ovarian fragments twelve patients were divided into small pieces in form cortex with medulla, randomly following four groups. Pieces Group 1 (n=30) operation, thawed their quality analyzed. 2 operation h, pre-cooling 5°C, 3 without pre-cooling, thawed, transplanted SCID mice then, culture 4 effectiveness pre-freezing cooling tissuewas evaluated by development follicles (histology) (FACS FITC-Annexin V Propidium Iodide). Results For groups 1, 2, mean densities per mm3 19.0, 20.2, 12.9, 12.2, respectively (P1-2, 3-4 >0.1). For these groups, 99%, 98%, 88% 90% preantral follicles, morphologically normal FACS analysis showed significantly decreased tissue (46.3% 33.6% Groups 4, respectively), contrast (77.1% 60.2 % 3, respectively, P1, 3-2, <0.05). Conclusions Long time (24 h) that evidences about increases viability thawing.