Effects of interactions between progesterone and prostaglandin on uterine contractility in a perfused swine uterus model.

BACKGROUND/AIM: Uterine quiescence at the time of embryo transfer is a prerequisite for successful in vitro fertilization (IVF). This study assessed whether prostaglandin-induced contractions in the perfused swine uterus can be reduced by progesterone. MATERIALS AND METHODS: Fifty-eight non-pregnant swine uteri were perfused using an established extracorporeal perfusion model. Intrauterine pressure changes during perfusion with prostaglandin (PG) administration (PGE1, PGE2, PGF2α) and progesterone (1 pg/ml, 10 pg/ml, 25 pg/ml, 50 pg/ml) were assessed using an intrauterine double-chip microcatheter. RESULTS: The contraction-stimulating effect of PGs was clearly reduced by progesterone. Only PGE1 still triggered relevant contractions during continuous perfusion with progesterone solution, up to a concentration of 10 pg/ml. With PGE2 and PGF2α, a clear reduction of uterine contractility was observed even at at a progesterone concentration of 1 pg/ml. CONCLUSION: The extracorporal perfusion model of swine uteri shows that PG-induced contractions can be reduced in a dose-dependent manner by progesterone.

Offspring after embryo-preserving biopsy of the embryoblast with standard ICSI equipment in mouse blastocysts.

BACKGROUND/AIM: Obtaining human embryonic stem cell lines has so far involved destroying the embryos. This has given rise to ethical concerns and is not permitted in most countries. This investigation tested whether removing multiple cells from blastocysts might allow continued embryonic development. MATERIALS AND METHODS: A total of 40 blastocysts from a black mouse strain were biopsied. The mouse blastocysts were fixed with a holding pipette. The zona pellucida and trophectoderm layer were penetrated with an injection pipette, and cells from the inner cell mass (ICM) were aspirated. The pipette was removed and the ICM cells were transferred into a medium. RESULTS: The blastocysts collapsed after pipette removal and were allowed to regenerate for 6 h. Twenty-four blastocysts recovered, expanded and were implanted into four white surrogate mothers. One surrogate mother gave birth to two black pups. CONCLUSION: This experiment demonstrates that nondestructive blastocyst biopsy from the ICM is possible in mice.

Uterus cryopreservation in the sheep: one step closer to uterus transplantation.

BACKGROUND: Uterus transplantation is the only way for women with no functional uterus to become pregnant. However, the technique is limited by the fact that an aggressive immunosuppression is necessary in order to avoid rejection of the graft. For better chances of finding a matching organ, which would minimize the immunosuppressive therapy, the establishment of cryobanks with a large number of uteri would be helpful. The aim of this study was to evaluate the feasibility to cryopreserve sheep uteri and to review the literature in this exciting new field of uterus cryopreservation and transplantation. MATERIAL AND METHODS: Ten sheep uteri were frozen either with slow cooling after perfusion with 10% dimethylsulfoxide, or without perfusion and the contractile ability was compared with that of fresh uteri. RESULTS: All perfused uteri showed contractions after thawing, similar to those of the non-frozen uteri. CONCLUSION: This study shows that the perfusion of sheep uterus with a cryoprotectant prior to slow freezing allows the cryopreservation of the whole organ and maintains the functionality of the organ after thawing. The perfused sheep uterus provides an experimental model for further investigations with other cryoprotective agents and freezing protocols.

Review. Fertility preservation for young female cancer patients.

Young female cancer patients are still being poorly counseled with regard to the negative impact of treatment on their fertility and on their options for fertility preservation. Today, many possibilities exist for fertility preservation, such as ovarian suppression with GnRH analogues, ovarian tissue cryopreservation, in vitro maturation or IVF after ovulation induction with aromatase inhibitors. A pregnancy after cancer treatment does not seem to limit the prognosis. This review focuses on both the effect of cancer treatments on fertility and on the various assisted-reproduction innovations that are available to provide the cancer patient with the option of future pregnancies. It is currently a time of uncertainty and revolution concerning the role of ovarian suppression and other fertility preservation measures in the management of early breast cancer, but developments in the near future promise to be very exciting.

Isolation of fibroblasts for coating of meshes for reconstructive surgery: differences between mesh types.

AIMS: An extensive colonization of surgical meshes with autologous fibroblasts may reduce complications. Therefore, we aimed to establish a technique that allows isolation and propagation of fibroblasts from vaginal biopsies. Using these cells we tested the applicability of several clinically applied meshes for fibroblast coating. MATERIALS & METHODS: Fibroblasts were isolated from vaginal tissue after digestion with collagenase. Characterization was performed by immunostaining for cytokeratin 5, 6 and 14, smooth muscle actin and vimentin. A semiquantitative technique was applied to determine the degree of mesh coating 5 h and 5 weeks after seeding of fibroblasts. Seven meshes of different mesh types have been tested. RESULTS: Cells with a fibroblast-like morphology have been isolated from vaginal tissue and could be propagated for at least 12 passages, resulting in a total number of 1.2 x 10(7) cells. Immunostaining showed that cells were positive for the mesenchymal cell marker vimentin and negative for smooth muscle actin, as well as the epithelial cell markers cytokeratin 5, 6 and 14, supporting their classification as fibroblasts. Clear differences in fibroblast colonization between the seven tested mesh types have been observed. Polypropylene mesh Obtape showed an acceptable covering with fibroblasts. The best coating was obtained for xenograft-based meshes, but under cell-culture conditions the mesh showed signs of decomposition. CONCLUSION: We have established a technique that allows isolation and propagation of vaginal fibroblasts. The result of vaginal fibroblast colonization of allograft-based meshes strongly depends on the mesh type, whereby the best coating could be achieved for a polypropylene mesh.

Pretreatment with gonadotropin-releasing hormone (GnRH) antagonists to prevent the flare-up effect of long-acting GnRH agonists: results of a pilot study.

This study evaluated in vivo whether the flare-up effect of GnRH agonists can be suppressed through pretreatment with a GnRH antagonist. The classic flare-up effect caused by 3.8 mg goserelin acetate could not be suppressed through pretreatment using a single dosage of 3.24 mg cetrorelix acetate.

Hormonal and histologic findings in human cryopreserved ovarian autografts.

This is the first report showing a hormonal and histologic discrepancy in cryopreserved human ovarian tissue 11 months after orthotopic autotransplantation. The presence of antral follicles was observed although the hormonal values had returned to castrated levels.

Differences in muscarinic-receptor agonist-, oxytocin-, and prostaglandin-induced uterine contractions.

OBJECTIVE: To investigate the contractile response of the perfused swine uterus to various receptor pathways (oxytocin, prostaglandins, and muscarine). DESIGN: An extracorporeal perfusion model of the swine uterus was used that keeps the uterus in a functional condition and is appropriate for the study of physiologic questions. INTERVENTION(S): Oxytocin-, prostaglandin-, and carbachol-induced uterine contractility and peristalsis were assessed using an intrauterine double-chip microcatheter. SETTING: University hospital. MAIN OUTCOME MEASURE(S): Intrauterine pressure profiles. RESULT(S): A dose-dependent increase in intrauterine pressure (IUP) in the isthmus uteri and corpus uteri was observed after the administration of prostaglandin F(2alpha) (PGF(2alpha)) and oxytocin, which reached a plateau after further stimulation. A dose-dependent increase in IUP in the isthmus uteri and corpus uteri was also observed after the administration of prostaglandin E(1) (PGE(1)) and prostaglandin E(2) (PGE(2)), with a plateau in IUP in the middle-concentration range and a decrease during the further course of stimulation. After administration of PGE(1), PGE(2), and PGF(2alpha), different directions of contraction waves were also observed. Carbachol also showed a unique contractility pattern, with isolated, very powerful, dose-dependent contractions with an IUP gradient, suggesting directed transport from the upper region to the lower region. CONCLUSION(S): This study demonstrated that oxytocin, the prostaglandins, and carbachol modulate contractility in nonpregnant swine uteri in a characteristic way, resulting in different contractility patterns.

Cancer and fertility preservation in females: where we stand and where we are heading.

It is estimated that, in 2010, one in every 250 adults will be a childhood cancer survivor. This review discusses the impact of current cancer treatment on fertility potential and the assisted-reproduction innovations available today for the most common cancers in young women. As the emerging discipline of fertility preservation is steadily attracting increasing interest, developments in the near future promise to be very exciting. However, in everyday routine work, better interdisciplinary cooperation between gynecological and pediatric oncologists, surgeons, immunologists and endocrinologists is necessary so that individualized options for fertility preservation can be offered in advance of surgical procedures or cancer treatments.

Cancer and fertility preservation: fertility preservation in breast cancer patients.

Aggressive chemotherapy has improved the life expectancy for reproductive-age women with breast cancer, but it often causes infertility or premature ovarian failure due to destruction of the ovarian reserve. Many questions concerning fertility preservation in breast cancer patients remain unanswered--for example, whether fertility preservation methods interfere with chemotherapy, and whether subsequent pregnancy has negative effects on the prognosis. Fertility preservation is a critical factor in decision-making for younger breast cancer patients, however, and clinicians should address this. The present article reviews the incidence of chemotherapy-induced amenorrhea, and discusses fertility-preservation options and the prognosis for patients who become pregnant after breast cancer.

Influence of resection volume on locoregional recurrence of breast cancer after breast-conserving surgery.

BACKGROUND: The goals of breast-conserving surgery are to provide the survival equivalent of mastectomy, a cosmetically acceptable breast and a low rate of locoregional recurrence in the treated breast. This retrospective study investigated the impact of the resection volume on locoregional recurrence after breast-conserving therapy in patients with early-stage invasive breast cancer. PATIENTS AND METHODS: Retrospective data from 185 women who were treated for operable breast tumours by breast-conserving surgery between 1995-1999 at the Martin-Luther-University in Halle/Germany were included in our study. Extent of total resection volume (TRV), tumour volume (V) and difference volume (DV) was compared for the influence on locoregional recurrence. RESULTS: Our data showed no significant correlation between the risk of locoregional recurrence and the extent of resection volume. Predictors of an increased risk of locoregional recurrence after breast-conserving surgery were large primary tumour, grading, lymphatic vascular invasion, hormone receptor status and lack of radiotherapy or hormonal therapy. CONCLUSION: According to the accuracy of locoregional disease control and maintenance of the breast's shape, our results support conservative surgery in early-stage breast cancer followed by radiotherapy and adjuvant systemic therapy.

The extracorporeal perfusion of the female pig detrusor as an experimental model for the study of bladder contractility.

AIMS: The objective of the study was to establish an experimental model for the extracorporeal perfusion of the pig detrusor. In order to validate this model we examined some biochemical parameters and determined the effect of carbachol on the contractility of perfused female pig bladders. METHODS: Twenty-six pig bladders were perfused with Krebs-Ringer bicarbonate-glucose buffer for a period up to 5 hr with the aim to preserve a viable organ, which would be responsive to contraction-inducing agents. The intravesical pressure of the bladder as well as the intraarterial pressure of the vesical arteries were recorded before and after administration of carbachol. RESULTS: The perfusate pH, lactate, partial carbon dioxide tension, and the ATP content in the perfused tissue, all indicators of tissue ischemia or cell necrosis, showed a good preservation of the organ for up to 5 hr. Carbachol was able to induce contractions of the prefilled bladder with a complete draining of the bladder throughout the whole perfusion period. CONCLUSIONS: We could demonstrate that this perfusion system was able to preserve the pig bladder in a functional condition, appropriate for the study of physiological questions.

The effect of a GnRH agonist on cryopreserved human ovarian grafts in severe combined immunodeficient mice.

This prospective study compares the effect of a GnRH agonist on the number of follicles in different developmental stages in cryopreserved human ovarian grafts transplanted into gonadotropin-stimulated or not stimulated severe combined immunodeficient mice (SCID mice). Human ovarian tissue from seven patients was cryopreserved with an open-freezing system and xenotransplanted in SCID mice. The SCID mice were then treated according to different stimulation protocols. The survival of the tissue after cryopreservation was examined by LIVE/DEAD viability staining or transplanted in the neck muscle of 41 SCID mice. Development of follicles, estradiol production, vaginal cytology, and uterus weight were assessed after 15 weeks with or without gonadotropin stimulation. Viable follicles were detected in all frozen/thawed specimens using the LIVE/DEAD assay. Triptorelin, a GnRH agonist, caused a significant reduction of follicles in all developmental stages in the non-gonadotropin-stimulated animals (P<0.001). In gonadotropin-stimulated animals, GnRH agonist treatment has no significant effect on primordial, primary and preantral follicle count, whereas the antral follicles were significantly fewer (P = 0.03). The GnRH agonist treatment is not able to prevent the primordial follicle depletion after the xenografting of ovarian tissue in SCID mice with or without gonadotropin stimulation. Furthermore, it causes an additional loss of follicles if administered during the critical neovascularization period after the transplantation.

Significant loss of primordial follicles after prolonged gonadotropin stimulation in xenografts of cryopreserved human ovarian tissue in severe combined immunodeficient mice.

This study evaluated the effect of gonadotropin stimulation on the primordial follicle reserve of cryopreserved human ovarian tissue after transplantation in severe combined immunodeficient mice. We found that prolonged gonadotropin stimulation significantly reduces primordial follicles.

The effect of cancer treatment on female fertility and strategies for preserving fertility.

Aggressive chemotherapy and radiotherapy in young patients with cancer has greatly enhanced the life expectancy of these patients, but these treatments often cause infertility because of the massive destruction of the ovarian reserve resulting in premature ovarian failure (POF). This review focuses on the effect of cancer treatments on fertility and on the various surgical and assisted-reproduction innovations that are available to provide the patient with the option of future pregnancies. As the emerging discipline of fertility preservation is steadily attracting increasing interest, developments in the near future promise to be very exciting. However, in everyday routine work, better interdisciplinary cooperation between gynecological and pediatric oncologists, surgeons, immunologists and endocrinologists is necessary so that individualized options for fertility preservation can be offered in advance of surgical procedures or cancer treatments. GnRH analog treatment can preserve fertility in some patients, but not in all. At present, cryopreservation of ovarian tissue appears as a very promising method of providing the cancer patient with a realistic chance of preserving fertility-a prospect that is also extremely important to patients for psychological reasons.

Reproduction beyond cancer: a message of hope for young women.

OBJECTIVE: Aggressive chemotherapy and radiotherapy or radical oncological surgery in young women with cancer has greatly enhanced these patients' life expectancy, but these treatments often cause infertility or premature ovarian failure due to a massive destruction of the ovarian reserve. The objective of this review is to discuss the effect of the various cancer treatments on fertility and present the various fertility sparing operations and fertility preservation strategies. METHOD: An extensive survey of the most up-to-date literature was performed. RESULTS: This review discusses the impact of current cancer treatment on fertility potential and the various surgical and assisted-reproduction innovations available today for the most common cancers in young women. Although the ability to retain reproductive potential is becoming a major quality-of-life factor in an increasing number of young female cancer survivors, they are still being poorly counseled with regard to the negative impact of the treatment on their fertility and on their options for fertility preservation. CONCLUSION: As the emerging discipline of fertility preservation is steadily attracting increasing interest, developments in the near future promise to be very exciting. However, in everyday routine work, better interdisciplinary cooperation between gynecological and pediatric oncologists, surgeons, immunologists, and endocrinologists is necessary so that individualized options for fertility preservation can be offered in advance of surgical procedures or cancer treatments.

Perfused non-pregnant swine uteri: a model for evaluating transport mechanisms to the side bearing the dominant follicle in humans.

Adequate uterine contractility and periovulatory peristalsis, interpreted as "rapid sperm transport" to the side bearing the dominant follicle, may be a precondition for successful reproduction in humans. Estrogen and progesterone fluctuate characteristically during the menstrual cycle, and their source is the dominant follicle and corpus luteum. The question is, how is the direction to the left or right side of transport mechanisms influenced? An extracorporeal perfusion model of the swine uterus was used that maintained the uterus in a functional condition and that was suitable for the study of physiological questions. The effects of side-dependent estrogen, progesterone, and estrogen plus progesterone perfusion on oxytocin-induced uterine peristalsis were assessed using two intrauterine microcatheters placed in each horn of the swine uterus. Estrogen perfusion was associated with an increase in intrauterine pressure (IUP) in a dose-dependent manner only in the estrogen-perfused horn of the swine uterus. There was a significant difference between the IUP increase measured in the estrogen-perfused horn and that in the non estrogen-perfused horn of the swine uterus. Progesterone perfusion showed no effect in general. Furthermore, progesterone antagonized the estrogen effects. This study demonstrates that side-dependent estrogen perfusion resulted in side-dependent contractility in the swine uterus perfusion system used. These observations show that estrogen stimulates uterine contractility in the estrogen-perfused uterine horn and that estrogens may be the "trigger" for the transport mechanisms to the side bearing the dominant follicle during the periovulatory phase through their locally increased concentration and distribution via the utero-ovarian counter-current system in humans.

Gonadal damage and options for fertility preservation in female and male cancer survivors.

It is estimated that in 2010, 1 in every 250 adults will be a childhood cancer survivor. Today, oncological surgery, radiotherapy and chemotherapy achieve relatively high rates of remission and long-term survival, yet are often detrimental to fertility. Quality of life is increasingly important to long-term survivors of cancer, and one of the major quality-of-life issues is the ability to produce and raise normal children. Developments in the near future in the emerging field of fertility preservation in cancer survivors promise to be very exciting. This article reviews the published literature, discusses the effects of cancer treatment on fertility and presents the options available today thanks to advances in assisted-reproduction technology for maintaining fertility in male and female patients undergoing this type of treatment. The various diagnostic methods of assessing the fertility potential and the efficacy of in vitro fertilization (IVF) after cancer treatment are also presented.

Comparison of two freezing protocols in an open freezing system for cryopreservation of rat ovarian tissue.

AIM: To compare two freezing protocols in an automatic open-vessel freezing system for cryopreservation of rat ovarian tissue. METHODS: Ovarian tissue was transplanted heterotopically into the neck muscle, either without cryopreservation (group 1, n = 6) or with cryopreservation after equilibration with 1.5 mol/L dimethyl sulfoxide and propanediol (protocol A, group 2, n = 6) or 1.5 mol/L ethyl glycol (protocol B, group 3, n = 6). The ovarian tissue was examined with LIVE/DEAD fluorescent viability staining and histologically after isotransplantation. RESULTS: The healthy follicular loss (intact oocyte and >50% granulosa cells alive) due to cryopreservation was 15.5% with protocol A and 12.2% with protocol B. Histological examination showed follicles in all developmental phases in all groups: group 1, 35.5 +/- 5.7/mm(2) (mean +/- SD); group 2, 16.0 +/- 5.0/mm(2); group 3, 17.3 +/- 5.7/mm(2). The differences between groups 1 and 2 and between groups 1 and 3 were significant (P < 0.001). The difference between groups 2 and 3 was not significant (P = 0.33). CONCLUSIONS: These results demonstrate that the use of an open freezing system with both freezing protocols allows cryopreservation of rat ovarian tissue with equally good survival rates.