An In Vitro Perfusion Method For Metabolic Studies On Human Postmenopausal Ovaries

Abstract

Five ovaries from four postmenopausal women undergoing bilateral oophorectomy for benign conditions were prepared for vascular perfusion for 1 to 5 hours in a recirculating system containing oxygenized (5% CO2 + 95% O2) synthethic bicarbonate buffered medium (Medium 199 + 4% bovine serum albumin + antibiotics). Two histologically normal ovaries released measurable amounts of progesterone into the medium. One ovary, containing a simple cyst, released progesterone into the perfusate, and this release was further increased after the addition of hCG at a concentration of 10 IU/ml. A perfused cystadenofibroma released cAMP, progesterone, estradiol and testosterone into the perfusion medium with an augmented release of cAMP, estradiol and testosterone after the addition of hCG. hCG also had a stimulating effect on the release of prostanoids from this tumour. Fluid from tumour cysts, obtained prior to perfusion, contained measurable levels of gonadotrophins, steroids and prostanoids. The isolated perfused ovary model may provide a tool for further exploration of endocrine and oncological aspects of the postmenopausal ovary.

Seven days ex vivo perfusion of whole ewe ovaries with follicular maturation and oocyte retrieval: towards the development of an alternative fertility preservation method

“…However, the relatively intact histomorphology of both the arterial and venous ovarian pedicles in both groups after the extended ovarian perfusion, together with the good cellular survival and minimal tissue damage is a significant achievement considering the long perfusion times explored. Our results are consistent with, and expand on, the previous short-time ovarian perfusion studies: 2 h of perfusion in ewes (Wallin et al 2009), 5 h in human (Abrahamsson et al 1990;Milenkovic et al 2011a), 12 h in rabbit (Lambertsen et al 1976;Hamada et al 1977Hamada et al , 1979Kobayashi et al 1981a) and 20 h in rat and mouse (Koos et al 1984;Sogn et al 1984;Brännström and Flaherty 1995). Our results support the findings of another study that used a complex perfusion medium supplemented with insulin, IGF-I and FSH that resulted in a successful 96-h long perfusion time (Maffei et al 2016).…”

“…This study demonstrated successful harvesting of oocytes and to our knowledge, this is the longest ex vivo perfusion time (7 days) described to date for ewe ovaries. A number of studies have been performed on ovarian tissue perfusion from rodents (Koos et al 1984;Sogn et al 1984;Brännström et al 1987;Brännström and Flaherty 1995), rabbit (Lambertsen et al 1976;Hamada et al 1977Hamada et al , 1979Kobayashi et al 1981a), sheep (Wallin et al 2009;Milenkovic et al 2011b;Maffei et al 2016) and also from human tissue (Stähler et al 1974;Abrahamsson et al 1990;Milenkovic et al 2011a;McLaughlin et al 2018). However, a complete follicular development in vitro with subsequent oocyte maturation and live offspring has only been successful in rodent models (Eppig and Schroeder 1989;O'Brien et al 2003).…”

Whole sheep ovary cryopreservation: evaluation of a slow freezing protocol with dimethylsulphoxide

“…This method has been used by us extensively for studies of ovarian physiology of freshly isolated ovaries from experimental animals [41] and human ovaries [42]. To assess the function of the ovary, we stimulated the ovaries with the adenylate cyclase stimulator forskolin.…”

In vitro ovarian tissue and organ culture: a review

“…The requirements of proper oxygen and carbon dioxide levels are satisfied by bubbling gas (5% CO 2 in oxygen) through the medium that is perfused into the ovary, and modifications to conditions can be made by altering the components of the medium or the rate of perfusion. Abrahamsson et al (50) perfused ovaries collected from women undergoing hysterectomy and oopherectomy for up to two hours to determine steroid production and responsiveness to hCG in post-menopausal ovaries. A similar technique, called perifusion, involves placing ovaries into incubation chambers in which fluid is pumped past the ovary rather than through the inherent blood vessels (51, 52, Figure 1C).…”