Meiotic and Developmental Competence in Mice Are Compromised Following Follicle Development In Vitro Using an Alginate-Based Culture System1

Summary The present study investigated the effects of c-type natriuretic peptide (CNP) on the development of murine preantral follicles during in vitro growth (IVG). Preantral follicles isolated from ovaries of Kunming mice were cultured in vitro. In the culture system, CNP was supplemented in the experimental groups and omitted in the control groups. In Experiment 1, CNP was only supplemented at the early stage and follicle development was evaluated. In Experiments 2 and 3, CNP was supplemented during the whole period of in vitro culture. In Experiment 2, follicle development and oocyte maturity were evaluated. In Experiment 3, follicle development and embryo cleavage after in vitro fertilization (IVF) were assessed. The results showed that in the control groups in all three experiments, granulosa cells migrated from within the follicle and the follicles could not reach the antral stage. In the experimental groups in all three experiments, no migration of granulosa cells was observed and follicle development was assessed as attaining the antral stage, which was significantly superior to that of the control group (P < 0.0001). Oocyte meiotic arrest was effectively maintained, hence giving good developmental competence. In conclusion, CNP supplementation in the culture system during IVG benefited the development of murine preantral follicles.

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Effects of IGF-I bioavailability on bovine preantral follicular development in vitro

Reproduction ◽

10.1530/rep-06-0382 ◽

2007 ◽

Vol 133 (6) ◽

pp. 1121-1128 ◽

Cited By ~ 55

Author(s):

Fiona H Thomas ◽

Bruce K Campbell ◽

David G Armstrong ◽

Evelyn E Telfer

Keyword(s):

Follicular Development ◽

Follicle Development ◽

Dependent Manner ◽

Igf Binding Proteins ◽

Preantral Follicles ◽

Follicle Diameter ◽

Igf I ◽

Igf Binding ◽

Development In Vitro

The aim of this study was to determine the effect of regulation of IGF-I bioavailability on preantral follicle development in vitro. Bovine preantral follicles were cultured for 6 days in serum-free medium with increasing doses of Long R3 (LR3) IGF-I (an analog with low affinity for IGF-binding proteins (IGFBPs)), or human recombinant IGF-I (hrIGF-I). Follicle diameter and estradiol production were measured every second day. On day 6, ratios of oocyte/follicle diameter and oocyte morphology were assessed by histological examination, and IGFBP-2 and -3 were detected by immunocytochemistry and in situ hybridization respectively. Both types of IGF-I increased follicle diameter in a dose-dependent manner (P < 0.05) and increased estradiol production over control levels (P < 0.05). However, follicles treated with LR3 IGF-I and the highest concentration of hrIGF-I (1000 ng/ml) had smaller oocyte/follicle ratios, and increased oocyte degeneration, compared with controls or follicles treated with physiological concentrations of hrIGF-I (P < 0.05). IGFBPs were detected in cultured preantral follicles, indicating a requirement for regulation of IGF bioavailability during the early stages of follicular development. Specifically, IGFBP-3 mRNA was found to be expressed in oocytes, and IGFBP-2 immunoreactivity was detected in oocytes and granulosa cells of cultured follicles. In summary, the regulation of IGF-I bioavailability by IGFBPs is necessary for the co-ordination of oocyte and follicle development in vitro.

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Ovarian cryopreservation strategies and the fine control of ovarian follicle development in vitro

Annals of the New York Academy of Sciences ◽

10.1111/j.1749-6632.2011.05967.x ◽

2011 ◽

Vol 1221 (1) ◽

pp. 40-46 ◽

Cited By ~ 3

Author(s):

Joshua Johnson ◽

Pasquale Patrizio

Keyword(s):

Ovarian Follicle ◽

Follicle Development ◽

Ovarian Follicle Development ◽

Fine Control ◽

Development In Vitro

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205.The effect of FSH concentration during IVM and gamete co-incubation length during IVF on the development of unstimulated prepubertal ewe oocytes

Reproduction Fertility and Development ◽

10.1071/srb04abs205 ◽

2004 ◽

Vol 16 (9) ◽

pp. 205 ◽

Cited By ~ 2

Author(s):

K. M. Morton ◽

W. M. C. Maxwell ◽

G. Evans

Keyword(s):

Blastocyst Stage ◽

Developmental Competence ◽

Blastocyst Formation ◽

In Vitro Development ◽

Oocyte Collection ◽

Chi Squared ◽

Development In Vitro ◽

Stage 1 ◽

Vitro Development

The developmental competence of prepubertal oocytes can be increased by the administration of gonadotrophins prior to oocyte collection (1); but this is not possible with abattoir-sourced oocytes, and modifications to the IVP system may increase in vitro development. Experiments were conducted to determine the effects of FSH concentration (10, 20 or 60 μg mL-1) during IVM (5 replicates) and gamete co-incubation length (short: 2-3 h, long: 18-20 h) during IVF (6 replicates) on subsequent embryonic development. For both experiments ovaries were collected from prepubertal lambs (16-24 weeks) slaughtered at an abattoir and embryos produced in vitro (1). Data were analysed by chi-squared test. Oocyte cleavage at 48 hours post-insemination (hpi) was higher for oocytes matured in medium containing 20 (60/77; 77.9%) and 60 (56/73; 76.7%) than 10 μg mL-1 (40/67; 59.7%) FSH. Blastocyst formation (% cultured oocytes) on Day 7 (Day 0 = IVF) was higher for oocytes matured with 20 (31/77; 40.3%) than 10 (16/67; 23.9%) or 60 μg mL-1 (20/73; 27.4%). Oocyte cleavage at 48 hpi was reduced for short (36/57; 63.2%) compared with long (49/55; 89.1%) co-incubation, although blastocyst formation (% cultured oocytes; Day 7) did not differ between groups (22/57; 38.6% and 23/55; 41.8%, respectively). These results demonstrate that increasing the FSH concentration above normal levels during IVM of prepubertal lamb oocytes improves development in vitro. Gamete co-incubation length did not influence the proportion of oocytes progressing to the blastocyst stage. (1) Morton et al. (2003) Proc. Soc. Reprod. Fert. P18.

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Correction to: Synergy of Paracrine Signaling During Early-Stage Mouse Ovarian Follicle Development In Vitro

Cellular and Molecular Bioengineering ◽

10.1007/s12195-018-0550-y ◽

2018 ◽

Vol 11 (6) ◽

pp. 537-537

Author(s):

Hong Zhou ◽

Joseph T. Decker ◽

Melissa M. Lemke ◽

Claire E. Tomaszewski ◽

Lonnie D. Shea ◽

...

Keyword(s):

Early Stage ◽

Ovarian Follicle ◽

Follicle Development ◽

Paracrine Signaling ◽

Ovarian Follicle Development ◽

Development In Vitro

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308 VERBASCOSIDE TREATMENT DURING IN VITRO MATURATION IMPROVES THE EMBRYO DEVELOPMENT OF PREPUBERTAL OVINE OOCYTES

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab308 ◽

2015 ◽

Vol 27 (1) ◽

pp. 243

Author(s):

M. E. Dell'Aquila ◽

F. Ariu ◽

N. A. Martino ◽

F. Minervini ◽

A. Cardinali ◽

...

Keyword(s):

Embryo Development ◽

In Vitro Maturation ◽

Redox Status ◽

Developmental Competence ◽

Cleavage Rate ◽

Chi Squared ◽

Development In Vitro ◽

Positive Effect ◽

Olive Mill

Verbascoside (VB), a bioactive polyphenol from olive mill wastewater with known antioxidant activity, was shown to act as a pro-oxidant molecule, by impairing energy/redox status and embryo developmental competence of prepubertal ovine oocytes when added at micromolar concentrations in a continuative 24-h in vitro maturation (IVM) exposure protocol (1). The aim of the present study was to determine whether a lower (nanomolar) VB concentration and a shorter exposure time (2 v. 24 h) during IVM may improve the maturation rates of prepubertal ovine oocytes and their subsequent embryonic development in vitro. Cumulus-oocyte complexes derived from the ovaries of slaughtered 1-mo-old prepubertal sheep oocytes underwent IVM in TCM 199 with 10% oestrus sheep serum, 0.1 IU mL–1 of FSH/LH, and 100 µM cysteamine, in 5% CO2 in air at 38.5°C for 24 h. Based on our previous results (Dell'Aquila et al. 2014 Biomed. Res. Int. 2014, 878062), VB was added in the IVM medium at 1.03 nM, and 2 incubation times (24 and 2 h) were tested. In the 2-h exposure group, after 2 h of exposure to VB, oocytes were washed and cultured up to 24 h without VB. A group of oocytes were cultured in absence of VB, as controls. Matured oocytes were fertilized with frozen-thawed ram semen in SOF medium for 22 h and zygotes were cultured in vitro for 8 days. Metaphase II (MII) cleavage and blastocyst rates were analysed by Chi-squared test. Embryo quality was evaluated by staining and total cell count of the blastocyst and analysis of variance (ANOVA) was applied. Differences were considered to be significant when P < 0.05. Compared to controls, VB treatment at the concentration of 1.03 nM and 24 h of exposure had no effect on MII rates (196/268, 73% v. 226/323, 70% MII/cultured oocytes; P > 0.05). However, this treatment allowed to obtain significantly higher rates of cleaved embryos/MII oocytes (156/196, 80% v. 165/226, 73%; respectively; P < 0.05), blastocyst yield/cleaved embryos (59/156, 38% v. 45/165, 27%, respectively; P < 0.05), and total blastocyst cell numbers (108.62 ± 19.87 v. 89.61 ± 26.32, respectively; P < 0.05) compared to control oocytes. The VB treatment at the same concentration but for 2 h induced only significantly higher cleavage rate (196/210, 93% v. 165/226, 73%; P < 0.05). In conclusion, our results showed that VB treatment at 1.03 nM during 24 h of IVM exerted a positive effect on in vitro embryo development of prepubertal ovine oocytes by increasing the blastocyst yield and their quality. The hypothesis that VB at nanomolar concentrations may improve cumulus-oocyte energy/redox status is under investigation.The authors acknowledge support by the Regione Autonoma della Sardegna (LR 7, Agosto 2007, no. 7, CRP-17602). The authors thank Dr D. Bebbere and L. Falchi, Dept. Veterinary Medicine, Sassari, for statistical analysis.

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174 DEVELOPMENT AND IN VITRO PRODUCTION OF PREPUBERTAL NELORE HEIFERS LONG-ACTING PROGESTERONE EXPOSED

Reproduction Fertility and Development ◽

10.1071/rdv28n2ab174 ◽

2016 ◽

Vol 28 (2) ◽

pp. 217

Author(s):

R. Corrêa ◽

J. R. Maio ◽

J. Garcia

Keyword(s):

Embryonic Development ◽

Developmental Competence ◽

In Vitro Production ◽

Young Females ◽

Animal Reproduction ◽

Development In Vitro ◽

Long Acting ◽

Von Mises ◽

Vitro Production

Ovum pick-up (OPU) and aspirated oocytes produced by in vitro production (IVP) are obtained at random stages of the oestrus cycle and are exposed to different concentrations of oestradiol, progesterone, LH and FSH. These factors may influence the oocyte developmental competence of in vitro embryos. The studies with young females aroused the interest of the investigated for decades and showed that one of the advantages working with young animals would be the amount of follicles that develop are best quality when comparing with pubertal animals. However some studies have shown a reduction in the competence of prepubertal oocytes can be partly attributed to the smaller size of the oocyte, differences in protein synthesis and energy metabolism, delayed migration of cytoplasmic organelles and reduced activity of some enzymes. The aim of this study was to evaluate the influence of injectable long-acting progesterone in embryonic development of prepubertal Nelore females. The OPU and treatments were carried out on the farm João Martins, in the county of Guatapará, São Paulo, Brazil, and laboratory stages of production in vitro embryo, the Department of Preventive Veterinary Medicine and Animal Reproduction, UNESP-Jaboticabal. They were select 21 heifers, Nelore, prepubertal of 127 animals. The selection was based on age, bodyweight, and absence of corpus luteum. The selected animals were aged 18 to 20 months and not pregnant with average bodyweight was 268 kg. The donor oocytes were divided into 3 experimental groups crossover design as follows: Group (P0, n = 21), animals in this group received 2 placebo oily solution applications (1 mL), interval of 7 days beginning 14 days (Day 14) before the first aspiration (Day 0); Group (P7; n = 21): the animals received a placebo solution oily application (1 mL), 14 days (Day 14) and progesterone (P4) injection (150 mg) 7 days (Day 7) before aspiration; Group (P14, n = 21), animals in this group received 2 injections P4 applications (150 mg) with an interval of 7 days, the first 14 days (Day 14) and the second 7 days (Day 7) before aspiration. There were a total of 3 OPU an interval of 28 days. After the first follicular aspiration groups were divided again so that all the animals go through all treatments. After confirming the homoscedasticity (BoxCox) and normal (Cramér-von Mises test) data, was conducted the analysis of variance (ANOVA). The Tukey test was used for comparisons of mean variables. The groups P0, P7, and P14 had an average of 4.04, 5.03, and 4.43 embryos produced by session. To assess embryonic development, it was observed that the treated groups (P7 and P14) and control (P0) produced a greater amount of expanded blastocyst, 3.40 ± 3.74, 2.57 ± 2.67 and 3.14 ± 3.41, respectively (P > 0.05). It was observed differences (P < 0.05) in the early blastocyst production in the treated groups produce a greater amount. The use of long-acting injectable progesterone improved did not delay embryo development in vitro but did not alter the production of embryos from prepubertal Nelore heifers.

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