Effect of exogenous lactoferrin and phosphoglycerate mutase 2 on the cryopreservation of wild ruminant epididymal/ejaculated sperm and testicular tissue.

Spermatozoa collected from the cauda epididymis of wild ruminants are more cryoresistant than are ejaculated spermatozoa. This work examines the effects of lactoferrin (LF) and phosphoglycerate mutase 2 (PGAM2), which are abundant in the epididymal sperm of wild ruminants, as additives in Iberian ibex and mouflon sperm extenders. In addition, LF was added to a vitrification medium to determine whether it also provided protection during the cryopreservation of testicular tissue.

Cooling rate modifies the location of aquaporin 3 in spermatozoa of sheep and goat.

The freeze-thawing process induces osmotic changes that may affect the membrane domain location of aquaporins' (AQP) in spermatozoa. Recent studies suggest that changes in AQP3 localization allows better sperm osmo-adaptation, improving the cryoresistance. Ultra-rapid freezing is an alternative cryopreservation technique that requires less equipment than conventional freezing, and it is faster, simpler and can be used in the field.

Immunolocalisation of aquaporins 3, 7, 9 and 10 in the epididymis of three wild ruminant species (Iberian ibex, mouflon and chamois) and sperm cryoresistance.

Context: In the epididymis, epithelial cells manage changes in the luminal environment for proper sperm maturation. Moreover, aquaglyceroporins, a subgroup of aquaporins (AQP), modulate the transport of water, glycerol and other small molecules in epithelial cells.

Aims: We aim to characterise the lining epithelium, quantify its cell composition and immunolocalise the aquaglyceroporins AQP3, AQP7, AQP9 and AQP10 alongside the epididymal ductus of three wild ruminant species, and to determine if species-specific differences could be associated with cauda sperm cryoresistance variations.

Variation of existence and location of aquaporin 3 in relation to cryoresistance of ram spermatozoa.

Introduction And Objective: Osmotic changes during the process of freeze-thawing involve changes in the location of aquaporins (AQPs) in membrane domains of spermatozoa. Some AQPs, like aquaporin 3 (AQP3), are linked to sperm cryotolerance in the porcine species. Conspicuous individual variability exists between rams and their ejaculates, which may be classified as displaying good freezability (GFE) or poor freezability (PFE), depending on several endogenous and environmental factors.

DNA integrity and viability of testicular cells from diverse wild species after slow freezing or vitrification.

Introduction And Objective: Cryopreservation of testicular tissues offers new possibilities to protect endangered species, genetically valuable individuals or even the fertility potential of prepubertal individuals who have died unexpectedly. However, the use of this technique still remains a challenge. In this study, slow freezing and vitrification of testicular tissue was investigated to find out which cryopreservation method could better preserve the viability and DNA integrity of testicular germ cells in diverse wild species.

Expression of Aquaglyceroporins in Spermatozoa from Wild Ruminants Is Influenced by Photoperiod and Thyroxine Concentrations.

This work identified the presence of AQPs in frozen-thawed sperm of wild ruminants and assessed the influence of the interaction between photoperiod and thyroxine on AQP expression, and on testosterone secretion. Thyroxine and melatonin were administered to ibexes. In a second experiment, performed in mouflons, circulating thyroxine was reduced via treatment with propylthiouracil (PTU), and an artificial long day (LD) photoperiod established.

Xenotransplantation by injection of a suspension of isolated preantral ovarian follicles and stroma cells under the kidney capsule of nude mice.

Objective: To develop and test a novel approach to xenotransplantation of isolated preantral follicles underneath the kidney capsule of immunodeficient mice.

Design: Prospective experimental animal study.

Setting: Academic research unit.

Preservation of fertility in young cancer patients: contribution of transmission electron microscopy.

During the last decade, new technologies in reproductive medicine have emerged to preserve the fertility of women whose gonadal function is threatened by premature menopause or gonadotoxic treatments. To offer an individualized approach to these patients, different experimental procedures are under investigation, including oocyte cryopreservation and cryopreservation and transplantation of ovarian tissue in the form of cortical fragments, whole ovary or isolated follicles. This review shows that transmission electron microscopy (TEM), combined with other in-vivo and in-vitro analysis techniques, is a valuable tool in the establishment of new experimental protocols to preserve female fertility.

Development of antral follicles after xenografting of isolated small human preantral follicles.

Small human pre-antral follicles can be enzymatically isolated from the surrounding stroma, and are able to survive after 7 days of xenografting. The aim of the present study was to assess the developmental capacity of enzymatically isolated human follicles after long-term xenografting to severe combined immunodeficient (SCID) mice. Ovarian biopsies were obtained from three women 26-29 years of age.

Chick embryo chorioallantoic membrane (CAM) model: a useful tool to study short-term transplantation of cryopreserved human ovarian tissue.

Objective: To investigate the chorioallantoic membrane (CAM) model for the study of short-term transplantation of frozen human ovarian tissue.

Design: Prospective study.

Setting: Academic research unit.

Autotransplantation of frozen-thawed ovarian tissue in a young woman: ultrastructure and viability of grafted tissue.

This is the first report of the presence of ultrastructurally normal primordial and primary follicles, 13 months after autotransplantation of frozen-thawed human ovarian tissue. The stroma contained numerous viable and ultrastructurally normal blood vessels but showed poor cellular density.

Cryopreservation and xenotransplantation of human ovarian tissue: an ultrastructural study.

Objective: To analyze the ultrastructure of human ovarian follicles after cryopreservation and short-term xenografting.

Design: Prospective experimental study.

Setting: Academic gynecology and anatomy research units.

Short-term transplantation of isolated human ovarian follicles and cortical tissue into nude mice.

This study was designed to evaluate follicular survival and growth after short-term transplantation of fresh isolated human follicles and ovarian cortical tissue to nude mice. Ovarian biopsies were obtained from nine women undergoing laparoscopy. Twelve nude mice were xenografted with an ovarian cortical fragment in the right ovarian bursa, and a clot containing isolated follicles in the left, for a period of 7 days.

Spermatogonial survival after cryopreservation and short-term orthotopic immature human cryptorchid testicular tissue grafting to immunodeficient mice.

Background: Fertility preservation has become an urgent clinical requisite for prepubertal male cancer patients undergoing gonadotoxic treatment. As these patients do not yet produce spermatozoa for freezing, only immature tissue is available for storage. We studied the survival and proliferative activity of spermatogonia and Sertoli cells after cryopreservation of cryptorchid testicular tissue pieces followed by xenografting for 21 days.

Apoptosis and ultrastructural assessment after cryopreservation of whole human ovaries with their vascular pedicle.

Objective: To investigate possible damage caused by freeze-thawing whole human ovaries.

Design: Prospective experimental study.

Setting: Academic gynecology research unit in a university hospital.

Laparoscopic ovariectomy for whole human ovary cryopreservation: technical aspects.

Objective: To describe the technique of laparoscopic ovariectomy with a view to cryopreservation of a whole ovary with its vascular pedicle.

Design: Descriptive study.

Setting: Gynecology research unit in a department of gynecology in a university hospital.

Ovarian tissue cryopreservation and transplantation: a review.

The review covers current options for ovarian tissue cryopreservation and transplantation and provides a systematic review of the existing literature from the last 10 years, taking into account all previously published reviews on the subject. The different cryopreservation options available for fertility preservation in cancer patients are embryo cryopreservation, oocyte cryopreservation and ovarian tissue cryopreservation. The choice depends on various parameters: the type and timing of chemotherapy, the type of cancer, the patient's age and the partner status.

Orthotopic transplantation of fresh ovarian cortex: a report of two cases.

Objective: To report two cases of orthotopic transplantation of fresh ovarian tissue.

Setting: Academic hospital.

Patient(s): Two patients with severe endometriosis, who underwent left oophorectomy for recurrent endometriosis.

Evaluation of Liberase, a purified enzyme blend, for the isolation of human primordial and primary ovarian follicles.

Background: The purpose of this study is to evaluate the effectiveness of a standardized mixture of purified enzymes (Liberase), for the isolation of human ovarian follicles.

Methods: This is an experimental prospective study. Ovarian biopsies were obtained from eight young women undergoing laparoscopy for benign gynaecological disease.

The role of cryopreservation for women prior to treatment of malignancy.

Purpose Of Review: The purpose of this review is to investigate recent advances in xenografting, as well as in orthotopic and heterotopic autotransplantation of human cryopreserved ovarian tissue.

Recent Findings: The first livebirth after orthotopic transplantation of cryopreserved ovarian tissue was reported recently. We discuss this case and other cases of reimplantation of cryopreserved ovarian tissue, bearing in mind that many questions remain.

Human endometrial epithelial cells (EEC) constitutively express more intercellular adhesion molecule (ICAM)-1 than endometrial stromal cells (ESC) in culture.

Problem: Intercellular adhesion molecule-1 (ICAM-1) is thought to play an important role in pathophysiological processes in endometrial tissue. The aim of this study was to quantify and compare the expression of ICAM-1 mRNA and protein in cultured endometrial epithelial cells (EEC) versus endometrial stromal cells (ESC).

Method Of Study: EEC and ESC were isolated from human endometrium and cultured.

Efficacy of in vitro fertilization after chemotherapy.

Objective: To evaluate if in vitro fertilization (IVF) with embryo cryopreservation can be proposed to patients immediately after one or two regimens of chemotherapy.

Design: Retrospective study.

Setting: Academic research center and IVF unit.