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PubMed2002

Effect of polyvinyl alcohol (PVA) concentration during vitrification of in vitro matured bovine oocytes.

Polyvinyl alcohol (PVA) was used as a substitute for serum in a vitrification solution for in vitro matured bovine oocytes. In vitro matured bovine oocytes were cryopreserved in various vitrification solutions (VS) supplemented with different concentrations (0.05, 0.1, 0.5, and 1%) of PVA, 20% fetal calf serum (FCS) or without macromolecule supplementation in a gel-loading tip (GL-tip). After warming, vitrified oocytes were examined for effects on survivability, fertilizability, and embryonic development in vitro. At 18 h in vitro fertilization after vitrifying and warming, the number of surviving mature oocytes vitrified in VS without macromolecule supplementation was significantly (P < 0.05) lower than those with macromolecule supplementation. For fertilizability after vitrification, there was no significant difference in the penetration rate of oocytes among fresh oocytes (98.7%); oocytes vitrified in VS supplemented with 0.1 (76.8%), 0.5 (70.2%), or 1% (80.3%) PVA; 20% (84.1%) FCS; or without supplementation (61.7%). Also, the normal fertilization rate was not significantly different in oocytes vitrified with 0.1 (56.5%), 0.5 (43.5%), or 1% (49.7%) PVA and 20% (60.6%) FCS, compared with fresh oocytes (84.0%). Subsequently, vitrified oocytes were examined for embryonic development effects in vitro. The highest proportion of cleaved oocytes after vitrification was obtained in VS supplemented with 0.1% (18.8%) PVA. Additionally, the proportion of development to morula stage (7.7%) in the oocytes vitrified in a VS supplemented with 0.1% PVA was significantly (P < 0.05) superior to that of the 0, 0.5, and 1% PVA-vitrified groups. However, the beneficial effect of PVA addition was not found in blastocyst development. Embryonic development of vitrified oocytes was significantly lower than that of fresh oocytes. In conclusion, the present results indicate that 0.1% PVA supplementation in VS results in a significantly higher rate of morula stage embryos than 0, 0.5, and 1% PVA supplementation, and could replace FCS in VS for vitrification of in vitro matured bovine oocytes.

Animals↗

PubMed2005

Pregnancies and births resulting from in vitro matured oocytes fertilized with testicular spermatozoa.

PURPOSE: In vitro maturation (IVM) of immature human oocytes is an attractive option for the treatment of infertility. Similarly, intracytoplasmic sperm injection (ICSI) followed by testicular fine needle aspiration (TEFNA) is an important treatment for primarily male-factor infertility. This report highlights the combination of these two advanced assisted reproduction techniques, namely IVM and fertilization with TEFNA-retrieved spermatozoa by ICSI to overcome both of male and female infertility problems. METHODS: Before immature oocyte retrieval (IOR), gonadotropin stimulation was given for 3 or 5 days. Following IVM, and mature oocytes were inseminated by ICSI followed by TEFNA. RESULTS: Four couples with five completed treatment cycles were performed, and total of 36 immature oocytes were retrieved. Following 36 to 48 h of culture, 32 (88.89%, 32/36) oocytes became mature. The mature oocytes were inseminated with TEFNA-retrieved sperm, and 18 (56.25%, 18/32) oocytes were fertilized normally following ICSI. Eleven embryos were transferred in five cycles and two pregnancies and two singleton births were achieved in two patients. CONCLUSIONS: This result demonstrates that the successful pregnancies and live births can be established from embryos produced from in vitro matured oocytes that fertilized with testicular sperm.

Adult↗

PubMed2001

Effects of growth hormone on in vitro maturation of germinal vesicle of human oocytes retrieved from small antral follicles.

PURPOSE: Evaluation of the effect of GH, administered in vivo, on in vitro maturation of human GV oocytes obtained from small follicles. METHODS: The study included 88 ICSI women stimulated with long agonist protocol. They were randomly allocated to two groups (Group 1, 44 women with 86 GV oocytes), were cotreated with GH (Group 2, no GH: 44 women with 67 GV oocytes). GV oocytes from small follicles were matured in vitro, injected, and their development observed. RESULTS: Resumption of meiosis was significantly higher in Group 1 compared to Group 2 (60 and 37.3% respectively). Significantly more M2 oocytes were obtained in Group 1 than in Group 2 (45.5 and 23.9% respectively). Fertilization rates were also significantly higher (51.9 and 18.8% respectively). No embryos were obtained in Group 2, whereas 21.1% of fertilized oocytes in Group 1 cleaved into class A embryos. CONCLUSION: In vivo administration of GH enhanced in vitro maturation and fertilization of human GV oocytes retrieved from small antral follicles.

Adult↗

PubMed1999

The influence of enucleation on the ultrastructure of in vitro matured and enucleated cattle oocytes.

The enucleation of recipient oocytes in nuclear transfer experiments is generally carried out by aspirating one third of the ooplasm adjacent to the first polar body. It was supposed that this enucleation step affects the ultrastructure of the remaining cytoplast, resulting in a decline or destruction of its cellular compartments. Even if the transferred nucleus had the potential to support the development of a single-cell nucleus transfer embryo to the blastocyst stage, meiotic division could be stopped at any stage if the destruction of the ultrastructure of host cytoplasm resulted in a limited metabolism. The present study was conducted to investigate the influence of the enucleation procedure on the ultrastructure of the remaining ooplast. In vitro matured oocytes; in vitro matured and enucleated oocytes; and in vitro matured and enucleated oocytes that were subsequently cultivated in vitro for additional 4 h were prepared for transmission electron microscopy (TEM). An examination of ultra-thin sections showed that the arrangement of organelles in all matured oocytes was in accordance with that already described for normal oocyte development. Immediately after enucleation no major differences in the arrangement of cortical granules, mitochondria, smooth endoplasmic reticulum (SER), lipid droplets and vacuoles were found compared with nonmanipulated oocytes. After enucleation and 4 h of culture, 24- and 36-h matured oocytes differed from each other in the arrangement of large aggregates of SER surrounded by a wall of mitochondria and lipid droplets. These complexes were still found in the 24-h but not in 36-h matured, enucleated and cultivated oocytes. Clusters of SER, mitochondria and lipid droplets were described by different authors as having metabolic activity. The results of this study in connection with results from nuclear transfer experiments suggest that these aggregates and their metabolic activity can be transferred with cytoplasm from 24- but not 36-h matured oocytes. Only cytoplasm from the 24-h matured oocytes showed a development-supporting effect when fused to enucleated recipient cells before nuclear transfer.

Animals↗

PubMed1992

The culture of human epididymal epithelium and in vitro maturation of epididymal spermatozoa.

OBJECTIVE: To promote human sperm maturation in vitro. DESIGN: Spermatozoa from the proximal epididymis were coincubated with epididymal epithelial fragments. SETTING: Hospital and research institute. PATIENTS, PARTICIPANTS: Tissue samples were obtained from men undergoing epididymovasostomy procedures or vasectomy. INTERVENTIONS: Fragments of epididymal epithelium formed everted epithelial spheres that in the presence of androgen maintained cell integrity. Coincubation for up to 48 hours of caput epididymal spermatozoa with 3-day-old epithelial cultures from the cauda epididymis was undertaken. MAIN OUTCOME MEASURES: Morphology of epididymal epithelium was assessed by light and electron microscopy. Pulse labeling of tissue in vitro with 35S-methionine was performed with analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique and fluorography. Spermatozoa were assessed for progressive motility and their capacity to bind to salt-stored human zona pellucidae. RESULTS: Epididymal fragments formed everted epithelial spheres that maintained cell integrity and functional morphology for 5 to 7 days. Specific proteins were synthesized in culture, in particular, proteins of 20, 22, 40, and 66 kd. Coincubation of caput epididymal spermatozoa with cultures from the cauda epididymis induced a significant increase in progressive sperm motility and sperm binding to salt-stored human zona pellucidae compared with control cultures of epithelium incubated in the absence of androgens or overgrown with fibroblasts. CONCLUSIONS: Aspects of human sperm maturation processes can be mimicked in vitro using coculture techniques with epididymal epithelium. This method may be valuable for improving the fertilizing capacity of human spermatozoa retrieved from the proximal region of the excurrent ducts.

Cells, Cultured↗

PubMed1999

Production and lambing rate of blastocysts derived from in vitro matured oocytes after gonadotropin treatment of prepubertal ewes.

The aim of this study was to evaluate the effect of gonadotropin treatment on the in vitro maturation, blastocyst production, and developmental potential to term of oocytes collected from Sardinian neonatal and prepubertal ewes at 4 to 6 wk of age. Cumulus-oocyte complexes were recovered at 24 h after withdrawal of a 1/6th size progestagenated pessary from the donors, of which each received 120 IU FSH/LH and 400 IU PMSG in a single dose 36 h before sponge removal. Treated donors produced a greater (P<.01) number of oocytes per animal (86.2 +/-7.9) compared with slaughterhouse (untreated) prepubertal ewes (55.5+/-6.1) of the same age or with treated neonatal ewes (6.1+/-0.7) 10 d old. During oocyte maturation, there were no differences in the percentage of germinal vesicle break-down (78.08 vs. 74.24), metaphase I (89.13 vs. 87.18), and metaphase II (77.91 vs. 76.38) when evaluated after 8, 14, and 24 h of maturation, respectively, between oocytes from treated and slaughterhouse (untreated) prepubertal ewes. The embryo cleavage (71.1 vs. 73.7) and blastocyst rates (22.2 vs. 19.8) were similar in the treated and the untreated prepubertal ewes after transfer of in vitro matured oocytes into ligated oviducts of temporary recipients. The in vitro viability rates of vitrified blastocysts (81.2 vs. 76.9) and the in vivo survival rates (46.1 vs. 50.0) of embryos derived from in vitro matured and in vivo fertilized oocytes showed no difference. The data suggest that gonadotropin treatment increases oocyte production per animal but has no effect on oocyte quality because embryo production and lambing rates of blastocysts derived from in vitro matured oocytes were not markedly different from those derived from untreated prepubertal ewes of the same age.

Animals↗

PubMed2001

A novel system for in vitro maturation of human oocytes.

OBJECTIVE: To compare in vitro maturation of cumulus-free oocytes in glucose-free medium (P1) and standard medium (TC199). DESIGN: Prospective, cohort study. SETTING: Assisted reproductive technology program. PATIENT(S): One hundred eight patients undergoing ICSI. INTERVENTION(S): Germinal vesicle-stage or metaphase I--stage oocytes were allocated to culture with P1 or TC199. Metaphase II oocytes were fixed for immunofluorescence analysis or fluorescence in situ hybridization at 24 or 48 hours (or both). Media were compared by performing conditional logistic regression analysis that controlled for egg-specific factors. MAIN OUTCOME MEASURE(S): Proportion of mature oocytes and appearance of normal spindle-chromosome cytoarchitecture. RESULT(S): At 24 hours, more P1 oocytes than TC199 oocytes reached metaphase II (59.7% vs. 44.9%). At 48 hours, 71.7% of P1 oocytes and 61.0% of TC199 oocytes reached metaphase II, but this difference was not significant. Metaphase II oocytes in P1 were 34.3% more likely than those in TC199 to have a bipolar spindle with aligned chromosomes. Compared with oocytes at the germinal vesicle stage at 0 hour, those at metaphase I at 0 hour were more likely to progress to metaphase II (72.6% vs. 46.1% at 24 hours; 84.1% vs. 60.6% at 48 hours). CONCLUSION(S): P1 is superior to TC199 for in vitro maturation of granulosa-free human oocytes.

Adult↗

PubMed2006

First successful pregnancy outcome after preimplantation genetic diagnosis for aneuploidy screening in embryos generated from natural-cycle in vitro fertilization combined with an in vitro maturation procedure.

OBJECTIVE: To report on the first successful pregnancy performing preimplantation genetic diagnosis (PGD) on embryos obtained after combining natural IVF with in vitro maturation procedure. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 35-year-old patient with polycystic ovaries who was attending the fertility clinic. INTERVENTION(S): In vitro maturation of immature oocytes and preimplantation genetic diagnosis. MAIN OUTCOME MEASURE(S): Preimplantation genetic diagnosis of aneuploidy screening in embryos produced from in vitro maturation procedure. RESULT(S): Thirteen immature and two mature oocytes were collected, and eight mature oocytes were normally fertilized, of which six embryos were biopsied for chromosome analysis. Two chromosomally normal embryos were transferred on day 5, resulting in the birth of a healthy child. CONCLUSION(S): This case report demonstrates that an acceptable number of embryos can be generated after in vitro maturation procedure to perform PGD for aneuploidy screening and that this approach can be extended to patients with polycystic ovaries or polycystic ovary syndrome who are undergoing PGD for other genetic diseases.

Aneuploidy↗

PubMed1983

On the effects of cyanoketone on gonadotrophin- and steroid-induced in vitro maturation of Oryzias oocytes.

Effects of cyanoketone (CK) on the gonadotrophin- and steroid-induced maturation in vitro of Oryzias latipes oocytes were examined. In oocyte covered with intact follicle or granulosa cell layers, CK inhibited gonadotrophin- and pregnenolone-induced maturation but not 20 beta-OH-progesterone-induced maturation. However, when follicle cells were removed completely from the oocytes, CK conspicuously lacked inhibitory effect on pregnenolone-induced oocyte maturation. It was ascertained by using purified pregnenolone that pregnenolone-induced maturation was not due to contamination with other maturation-inducing steroids. It was also observed immunocytochemically that a purified pregnenolone stimulated in vitro production of progestins in the granulosa cells, and that CK inhibited this stimulation. These results indicate that pregnenolone, like gonadotrophin, induces maturation of intrafollicular oocytes of O. latipes by stimulation of production of such steroids as progesterone in follicle cells (granulosa cells), while it can directly induce maturation of oocytes not surrounded by follicle cells.

Androstenols↗

PubMed1996

Effects of injecting calcium chloride into in vitro-matured porcine oocytes.

In vitro-matured porcine oocytes were given injections of 0.1 M CaCl2 and after 6 h evaluated for signs of early and late activation events. CaCl2 injection caused cortical granule exocytosis in 75% (3 of 4) of the oocytes tested. It also induced cell cycle resumption as monitored by the histone H1 kinase assay: the phosphorylation rate of histone H1 decreased to 36.7% of the original value. Treated oocytes completed meiosis, extruded the second polar body, and progressed to first interphase: 79.4% of them formed one or more pronuclei. The elevated intracellular Ca2+ level resulted in activation-related changes in the protein synthetic profile in 90% (9 of 10) of the oocytes. Furthermore, 14.7% (9 of 61) of the treated oocytes developed to the compact morula/early blastocyst stage after a 7-day culture in ligated porcine oviduct, and one blastocyst hatched from the zona pellucida. Control oocytes given injections of 0.1 M MgCl2 or carrier medium (10 mM Hepes) did not show the changes mentioned. The results strengthen the idea that Ca2+ is a cell messenger that plays a central part in oocyte activation; it is concluded that elevated intracellular Ca2+ level caused by a single injection of CaCl2 leads to both early and late events of porcine oocyte activation.

Animals↗

PubMed2005

Kinetics of nuclear maturation and effect of holding ovaries at room temperature on in vitro maturation of camel (Camelus dromedarius) oocytes.

Experiments were conducted to investigate kinetics of in vitro nuclear maturation and the effect of storing ovaries at room temperature on initial chromatin configuration and in vitro maturation of dromedary camel oocytes. Cumulus oocyte complexes (COCs) were collected from slaughterhouse ovaries and matured in vitro for 4-48h. At every 4h interval (starting from 0 to 48 h), groups of oocytes were fixed, stained and evaluated for the status of nuclear chromatin. Oocytes were categorized as germinal vesicle (GV), diakinesis (DK), metaphase-I (M-I), anaphase-I (A-I), metaphase-II (M-II) stage and those with degenerated, fragmented, activated or without a visible chromatin as others. At the start of culture, 74% (66/89) oocytes were at GV stage, 13% (12/89) at DK and 12% (11/89) were classified as others. Germinal vesicle breakdown started spontaneously in culture and at 20 h of culture 97% oocytes had already completed this process. After 8 and 16 h of maturation the highest proportion of oocytes (42%, 48/114 and 41%, 51/123) were at DK and M-I stage, respectively. The proportions of oocytes reaching M-II stage at 32 (42%, 50/118), 36 (45%, 47/104), 40 (49%, 57/117), 44 (52%, 103/198) and 48 h (46%, 55/120) of culture were not different from each other (P>0.05). The proportion of oocytes categorized as others, however, increased after 40 h of culture and was higher (P<0.05) at 48 h compared with other maturation periods. There was no difference (P>0.05) in the proportion of oocytes reaching M-II stage from the ovaries collected and stored in normal saline solution (NSS) at room temperature for 12h (43%, 64/148) and those collected in warm NSS (37 degrees C) and processed immediately after arrival in laboratory (49%, 57/117). However, low number of oocytes reached M-II stage from ovaries collected in warm NSS but stored at room temperature (29%, 37/128) compared with other two groups (P<0.05). It may be concluded that dromedary oocytes require 32-44h of in vitro culture to have an optimum number of oocytes in M-II stage. However, further studies are required to find out the most appropriate maturation period, which will result in the further development of these oocytes after IVF, ICSI, parthenogenetic activation or nuclear transfer. Ovaries can be collected and stored in normal saline solution at room temperature for 12h without any appreciable effect on the nuclear maturation of the oocytes.

Animals↗

PubMed2002

In vitro maturation of bitch oocytes from advanced preantral follicles in synthetic oviduct fluid medium: serum is not essential.

The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210 microm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n = 230); (2) SOF + 3 mg/ml bovine serum albumin (+BSA, n = 220); (3) SOF + 20% fetal bovine serum (+FBS, n = 227); or (4) SOF + 3 mg/ml BSA + 20% FBS (+BSA+FBS, n = 232), then cultured for up to 72 h. A group of control follicles was not cultured (n = 103). The percentages of oocytes reaching metaphase I to metaphase II stages (MI to MII) did not differ between treatments at each culture period. Within treatments, the percentages of oocytes at MI to MII stages did not differ with duration of culture. However, when compared to the control group (0.97%) the percentages of oocytes at MI to MII increased (P < 0.05) in the SOF group after 48 h (10.0%) and 72 h (12.2%) of culture. In the +BSA (10.1%) and +FBS (9.7%) groups, the percentages of oocytes at MI to MII increased (P < 0.05) above control values only after 72 h of culture. The percentage of oocytes at MI to MII did not significantly increase in the +BSA+FBS group (3.9,6.6 and 7.6% at 24,48 and 72 h of culture, respectively) compared to the control group. These results indicate that under the described conditions supplementation of culture medium with BSA or FBS is not essential, and the simple medium SOF can support nuclear maturation of a small proportion of bitch oocytes in vitro.

Animals↗

PubMed2002

Localisation of the hyaluronan receptor CD44 in porcine cumulus cells during in vivo and in vitro maturation.

Polyspermy is fairly common during porcine in vitro fertilisation (IVF), perhaps due to incomplete in vitro oocyte maturation (IVM). Porcine cumulus cells (CCs) layered around the oocyte produce large amounts of extracellular hyaluronan (HA) when forming an expanding cell cloud during the last phase of oocyte maturation. The specific actions of HA are mediated via HA-binding proteins (HABPs), such as CD44, which act as receptors. In this study using immunocytochemistry and western blotting we investigated the localisation of CD44 in CCs obtained from in vivo-matured pig cumulus-oocyte complexes (COCs) and compared it with that in CCs from immature COCs and of COCs subjected to IVM and IVF procedures. Immunolabelling of CD44 was absent or very weak in CCs from immature COCs but strongly present on the surface of the CCs obtained from in vivo, displaying a similar localisation in the in vitro-matured COCs. In the latter, the labelling decreased but did not disappear in CCs 4 h after sperm co-incubation during IVF. Immunoblotting detected bands of between 73 and 88 kDa, corresponding to CD44, in the protein extract from in vivo CCs collected immediately prior to, or following spontaneous ovulation. The in vitro-matured CCs, however, presented bands ranging from 81 kDa to 88 kDa. Also, the bands found in the in vivo-matured CCs showed a larger variation of intensity and migration among animals than did the batches of in vitro-matured CCs. No CD44 band was detected on aliquots of the frozen-thawed boar spermatozoa used for IVF. The results clearly demonstrate that the specific HA receptor CD44 is present in expanding CCs of in vivo-matured pig COCs, in relation to increasing amounts of inter-CC HA. The subtle differences in molecular weight and migration ability observed between in vivo and in vitro samples may relate to differences in glycosylation and thus explain differences in HA-binding ability, of consequence for optimising in vitro culture conditions.

Animals↗

PubMedDate not supplied

[The action of neurotransmitters and their antagonists on oocyte maturation. The action of serotonin antagonists on the in-vitro maturation of amphibian oocytes].

Serotonin antagonists (inmecarb hydrochloride and inmecarb methiodide) stimulate in vitro maturation of Bufo viridis and Xenopus laevis oocytes or potentiate the action of progesterone. In contrast to that, serotonin inhibits or blocks oocyte maturation stimulated by progesterone or serotonin antagonists. Sensitivity of B. viridis oocytes to serotonin, its agonists and antagonists is subject to seasonal changes. A maximum sensitivity of intact oocytes is observed in February-March, and that of denuded ones in May-June. We suggest that endogenous serotonin is involved in maintenance of the block of meiosis and in the oocyte maturation control in amphibians.

Animals↗

PubMed1996

Pregnancy and birth after intracytoplasmic sperm injection of in vitro matured germinal-vesicle stage oocytes: case report.

OBJECTIVE: To report a normal pregnancy and the delivery of a healthy child after the combination of in vitro maturation of germinal-vesicle stage oocytes and intracytoplasmic sperm injection (ICSI) in a patient. SETTING: Procedures were performed in a tertiary IVF center coupled with an institutional research environment. MAIN OUTCOME MEASURES: Maturation rate of immature oocytes after in vitro maturation and intactness, fertilization, and developmental rates of oocytes after microinjection. RESULTS: Nine of 14 germinal-vesicle stage oocytes matured to the metaphase II stage after 30 hours of in vitro culture (64%). Seven of eight injected and intact oocytes fertilized normally (78%) and five of them cleaved with < 20% fragmentation (71%). Four embryos were transferred and a singleton pregnancy was obtained that ended in the delivery of a healthy child. CONCLUSION: In vitro maturation of immature oocytes together with ICSI can result in normal fertilization, embryo development, pregnancy, and the delivery of healthy child.

Adult↗

PubMed2003

[In vitro maturation, fertilization and embryo transfer of human immature oocyte].

OBJECTIVE: To try a new method for treating infertile women by in vitro maturation technique. METHODS: To set up in vitro maturation (IVM) program, including stimulating regimen, oocyte retrieval technique, and culture system of the immature oocytes with the medium consisted of essential supplements and human mature follicular fluid. IVM was performed in 30 infertile patients (35 cycles), (1) with polycystic ovary syndrome (POCS) (14/30), (2) ovary hyperstimulation syndrome (OHSS) in the previous stimulating cycles (6/30), and (3) retardation of the oocyte growth in conventional IVF cycles (10/30). RESULTS: Totally 203 oocytes were obtained, with a mean of 5.8 oocytes each cycle. First polar bodies were observed in 156 oocytes, maturation rate was 76.8%. There were 120 oocytes getting into pronucleus stage, normal fertilization rate was 76.9%. Embryo transfer was performed in 33 cycles. Eight pregnancies were established, pregnancy rate was 24%. Five patients have delivered 7 healthy babies. CONCLUSIONS: The result of our study indicated that IVM is one of the new assisted reproductive technologies for the infertile women with PCOS, poor response to stimulation and the history of OHSS. Human mature follicular fluid would be an ideal component in the maturation medium for oocyte maturation in vitro and provides a much simplified and practical method for clinical use.

Adult↗

PubMed2003

In vitro maturation and fertilization of buffalo oocytes: effects of storage of ovaries, IVM temperatures, storage of processed sperm and fertilization media.

Studies were conducted to examine the possibility of preserving slaughterhouse-derived buffalo ovaries at 4 degrees C for 0 (control), 12 and 24 h to maintain the developmental competence of the oocytes (experiment 1), to assess the effect of incubation temperature during oocyte maturation on rates of in vitro maturation (IVM) and in vitro fertilization (IVF) of buffalo oocytes and embryo development (experiment 2), and to examine the effect of storage at 25 degrees C for 0 (control), 4 and 8 h of frozen-thawed buffalo sperm and BO and H-TALP as sperm processing and fertilization media on cleavage and embryo development in vitro of buffalo oocytes (experiment 3) in order to optimize the IVF technology in buffalo. Results suggested that storage of ovaries at 4 degrees C for 12 or 24 h significantly (p < 0.05) reduced the developmental potential of oocytes. Incubation temperatures during the IVM influenced the fertilization rate but had no significant effect on maturation and subsequent embryo development. The incubation temperature of 38.5 degrees C during IVM was found to be optimum for embryo production in vitro. Storage of frozen-thawed sperm at 25 degrees C for 8 h significantly (p < 0.05) decreased its ability to cleave the oocytes. Sperm processed in BO medium had significantly (p < 0.05) higher ability to cleave the oocytes than the H-TALP medium.

Animals↗

PubMed2006

In-vitro maturation of oocytes: biological aspects.

Natural cycle and in-vitro maturation (IVM) of oocytes are becoming interesting alternatives to classical assisted reproduction technology approaches for patients, especially in those at high risk for ovarian hyperstimulation syndrome or with poor ovarian reserve. More than for their clinical and biological indications, natural cycle and IVM of oocytes can also be considered as good social and economic alternatives to the classical IVF treatment, based on their financial cost-effectiveness with exclusion of expensive medications. To be successful, IVM must entail both nuclear and cytoplasmic maturation, and its maturation and success rates are affected by the number of collected cumulus layers, the degree of atresia and the maturation rate between 24 and 48 h. Endogenous regulation of oocyte maturation is a complex sequence of events regulated by endocrine parameters, oocyte/follicular cross-talk, and intra-oocyte kinase/phosphatase interactions. This complex process requires a better definition of each contributing factor affecting oocyte development and the resulting embryo quality. The clinical aspects of IVM have been documented earlier; the present paper will mainly focus on the biological aspect of oocyte maturation in vitro and the quality of derived embryos.

Animals↗