The best source of isolated stromal cells for the artificial ovary: medulla or cortex, cryopreserved or fresh?

Todorov

et al. 2016

Reprod Biol Endocrinol

BackgroundPhosphatidylserine is the phospholipid component which plays a key role in cell cycle signaling, specifically in regards to necrosis and apoptosis. When a cell affected by some negative factors, phosphatidylserine is no longer restricted to the intracellular side of membrane and can be translocated to the extracellular surface of the cell. Cryopreservation can induce translocation of phosphatidylserine in response to hypoxia, increasing intracellular Ca2+, osmotic disruption of cellular membranes, generation of reactive oxygen species and lipid peroxidation. As such the aim of this study was to test the level of phosphatidylserine translocation in frozen human medulla-contained and medulla-free ovarian tissue fragments.MethodsOvarian fragments from twelve patients were divided into small pieces of two types, medulla-free cortex (Group 1, n = 42, 1.5–3.0 × 1.5–3.0 × 0.5–0.8 mm) and cortex with medulla (Group 2, n = 42, 1.5–3.0 × 1.5–3.0 × 1.5–2.0 mm), pre-cooled after operative removal to 5 °C for 24 h and then conventionally frozen with 6 % dimethyl sulfoxide, 6 % ethylene glycol and 0.15 M sucrose in standard 5-ml cryo-vials. After thawing at +100 °C and step-wise removal of cryoprotectants in 0.5 M sucrose, ovarian pieces were xenografted to SCID mice for 45 days. The efficacy of tissues cryopreservation, taking into account the presence or absence of medulla, was evaluated by the development of follicles (histology with hematoxylin-eosin) and through the intensity of translocation of phosphatidylserine (FACS with FITC-Annexin V and Propidium Iodide).ResultsFor Groups 1 and 2, the mean densities of follicles per 1 mm3 were 9.8, and 9.0, respectively. In these groups, 90 and 90 % preantral follicles appeared morphologically normal. However, FACS analysis showed a significantly decreased intensity of translocation of phosphatidylserine (FITC-Annexin V positive) after cryopreservation of tissue with medulla (Group 2, 59.6 %), in contrast with tissue frozen without medulla (Group 1, 78.0 %, P < 0.05). In Groups 1 and 2 it was detected that 21.6 and 40.0 % cells were viable (FITC-Annexin V negative, Propidium Iodide negative).ConclusionThe presence of medulla in ovarian pieces is beneficial for post-thaw development of cryopreserved human ovarian tissue.

“…The aim of this study was to determine the best source of isolated ovarian cells for the artificial ovary: cortex or medulla [46]. …”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Cryopreservation and xenografting of human ovarian fragments: medulla decreases the phosphatidylserine translocation rate

Todorov

et al. 2016

Reprod Biol Endocrinol

Expert Review of Anticancer Therapy

“…In strategy three, oophropexy and pelvic shielding do not provide ovarian protection when chemotherapy is used, while GnRH analogs do not provide ovarian protection when radiotherapy is used [31, 32, 33, 34]. In order to avoid such different contraindications, new advances in research to restore fertility of female cancer patients have been recently attempted including artificial ovary [35, 36, 37, 38], and use of mesenchymal stem cells [39, 40, 41]. …”

Section: Expert Commentarymentioning

confidence: 99%

Anticancer treatments and female fertility: clinical concerns and role of oncologists in oncofertility practice

Salama

Woodruff

2017

Introduction: Anticancer treatments such as aggressive chemotherapy and radiotherapy have deleterious gonadotoxic side effects and are considered the most common causes of pathological and iatrogenic fertility loss in women. Areas Covered: In order to preserve fertility of young women and girls with cancer, several established, experimental, and debatable options can be offered in the emerging field of oncofertility. This article reviews the effects of anticancer treatments on female fertility and discusses the current challenges and future directions of fertility preservation options that can be offered to the female patients with cancer. Expert Commentary: Although promising, several medical, economic, social and legal barriers face oncofertility practice around the globe especially in underserved areas. To overcome such barriers, more effective solutions should be provided to spread awareness and enhance communication between patients, oncologists and gynecologists. Early referral by oncologists before initiation of chemotherapy and radiotherapy is an important key factor for success in female fertility preservation strategies.

Gonadal Tissue Cryopreservation in Fertility Preservation

“…Soares et al [53] evaluated the best source of ovarian cells to be grafted inside the artificial ovary: cortex or medulla, cryopreserved or fresh. Their findings suggest that ovarian cells from fresh medullary tissue can be isolated in larger numbers, show higher viability, and are able to improve graft vascularization.…”

Section: Cell Populationmentioning

confidence: 99%

Artificial Ovary

Amorim¹

2016

Survival rates of many malignant diseases are steadily improving, but for patients of childbearing age, fertility restoration often becomes a vital concern after disease remission. In women, treatments such as chemo/radiotherapy can be very harmful to the ovaries, causing loss of both endocrine and reproductive functions. When gonadotoxic treatment cannot be delayed, ovarian tissue cryobanking is the only way of preserving fertility. However, this technique is not advisable for patients with certain types of cancer, since there is a risk of reintroducing malignant cells present in the cryopreserved tissue. For these patients, a safer alternative could be transplantation of isolated preantral follicles back to their natural environment. To encapsulate and protect isolated follicles, a transplantable artificial ovary needs to be created. The main goal of the artificial ovary is to mimic the natural organ, and for this, it should be composed of a matrix that encapsulates and protects not only the isolated follicles but also autologous ovarian cells and bioactive factors, which are necessary for follicle survival and development. The aim of this chapter is to describe this new technology, its indications, advantages, and the different approaches to create it.