結合慢速冷凍+快速解凍，可能比傳統玻璃化冷凍+快速解凍
更有效保存卵巢組織
傳統玻璃化冷凍+快速解凍下，卵細胞粒線體之受損率，卵巢組織內部粒線體受損率明顯高於外部之受損率‧說明冷凍前需要足夠時間使其抗凍液作用於卵巢組織‧
Hum Reprod. 2016 Jun 9. pii: dew134. [Epub ahead of print]
Morphological, ultrastructural and functional imaging of frozen/thawed and vitrified/warmed human ovarian tissue retrieved from oncological patients.
Abstract
STUDY QUESTION:
Which is the best method for human ovarian tissue cryopreservation: slow freezing/rapid thawing (SF/RT) orvitrification/warming (V/W)?
SUMMARY ANSWER:
The conventional SF/RT protocol used in this study seems to better preserve the morpho-functional status of human cryopreserved ovarian tissue than the used open carrier V/W protocol.
WHAT IS KNOWN ALREADY:
Cryopreservation of human ovarian tissue is generally performed using the SF/RT method. However, reduction in the follicular pool and stroma damage are often observed. An emerging alternative procedure is represented by V/W which seems to allow the maintenance of the morphological integrity of the stroma.
STUDY DESIGN, SIZE, DURATION:
This is a retrospective cohort study including six patients affected by oncological diseases and enrolled from January to December 2014.
PARTICIPANTS/MATERIALS, SETTING, METHODS:
Ovarian tissue was laparoscopically harvested from the right and left ovaries and was cryopreserved using a routinary SF/RT protocol or a V/W method, involving tissue incubation in two solutions (containing propylene glycol, ethylene glycol and sucrose at different concentrations) and vitrification in an open system. For each patient, three pieces from each ovary were collected at the time of laparoscopy (fresh tissue) and after storage (SF/RT or V/W) and processed for light microscopy (LM) and transmission electron microscopy (TEM), to assess the morphological and ultrastructural features of follicles and stroma, and for laser scanning confocal microscopy (LSCM), to determine the functional energetic/redox stroma status. The preservation status of SF/RT and V/W ovarian tissues was compared with that of fresh ones, as well as between them.
MAIN RESULTS AND THE ROLE OF CHANCE:
By LM and TEM, SF/RT and V/W samples showed cryodamage of small entity. Interstitial oedema and increased stromal cell vacuolization and chromatin clumping were observed in SF/RT samples; in contrast, V/W samples showed oocyte nuclei with slightly thickened chromatin and irregular shapes. The functional imaging analysis by LSCM revealed that the mitochondrial activity and intracellular reactive oxygen species levels were reduced both in SF/RT and in V/W samples compared with fresh samples. The study also showed progressive dysfunction of the mitochondrial activity going from the outer to the inner serial section of the ovarian cortex. The reduction of mitochondrial activity of V/W samples compared with fresh samples was significantly higher in the inner section than in the outer section.
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