Improvement of the in vitro growth and maturation of isolated mouse preantral follicles in the presence of repaglinide

2019 ◽  
Vol 9 (o3) ◽  
Author(s):  
Sura A. Awadh ◽  
Mehri Azadbakht ◽  
Faris N. A. Alhady
Keyword(s):  
Oocyte Maturation ◽  
Ovarian Follicle ◽  
Control Group ◽  
In Vitro Growth ◽  
Preantral Follicles ◽  
Maturation Rate ◽  
Reproductive Techniques ◽  
In Vitro Oocyte Maturation ◽  
Survival Density

The development of in vitro culture systems that result to preantral follicles growth and increasing of developmental competency of oocytes obtained from follicles has an important role in fertility preservation and assisted reproductive techniques. In this research, we evaluated the effect of repaglinide on in vitro growth and maturation of preantral follicles. Preantral follicles were isolated from 12-14 day-old female NMRI mice ovaries and cultured for 12 days cultured in α-MEM (Control), α-MEM supplemented with 1µM of repaglinide. Follicles examined for development on 1, 3, 6, 9, 12 days of culture. At the end of culture period after HCG administration in vitro oocyte maturation was assessed. Results showed that in vitro follicle growth, survival, density of granulosa cells and steroidogenic activity were higher than the control group (p less than 0.05). In vitro maturation rate in oocytes derived from follicles in the treatment group was higher than control group (p less than 0.05). Therefore the supplementation of the culture medium with repaglinide can improve the ovarian follicle survival, growth and subsequently in vitro oocyte maturation.

scholarly journals Pengaruh urea dalam media maturasi in vitro terhadap tingkat maturasi oosit sapi

2021 ◽  
Vol 10 (2) ◽  
pp. 46
Author(s):  
Sepvian Dewi Kurniawati ◽  
Suryanie Sarudji ◽  
Widjiati Widjiati
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Metaphase Plate ◽  
Petri Dish ◽  
Control Group ◽  
Maturation Medium ◽  
First Polar Body ◽  
Maturation Rate ◽  
Follicle Aspiration

This study was aimed to determine the effect of urea in maturation medium on in vitro oocyte maturation rate. The medium used was TCM-199 added with Hepes, NaHCO3, Kanamycin 0.15 IU/mL, PMSG, 0.15 IU/mL hCG, and 10% FBS. Cumulus oocyte complexes (COCs) of cows derived from follicle aspiration were divided into three groups. In control group (P0), the COCs were matured in vitro in a maturation medium without urea addition, meanwhile in the P1 and P2 groups, the medium was added with urea 20 and 40 mg/dL, respectively. Each petri dish contained three drops of maturation medium (300 µl/drops) according to the groups. Microdrops were coated with mineral oil and then incubated in a 5% CO2 incubator, at 39 ˚C with maximum humidity. Aceto-orcein staining was conducted to evaluate the maturation of oocytes based on the achievement of metaphase II phase that is indicated by the presence of metaphase plate and/or first polar body. The result showed that the oocyte maturation rates of P0, P1, and P2 were 51.25, 52.43 (p >0.05), and 46.88 % (p <0.05) respectively. It could be concluded that the presence of urea at 40 mg/dL in maturation medium reduced the percentage of bovine oocyte maturation in vitro.

scholarly journals Use of Melatonin in the In Vitro Production of Bovine Embryos

2020 ◽  
Vol 21 ◽  
Author(s):  
Alan da Silva LIRA ◽  
Ricardo de Macedo CHAVES ◽  
Felipe de Jesus MORAES JUNIOR ◽  
Sergio Henrique COSTA JUNIOR ◽  
Brenda Karine Lima do AMARAL ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Control Group ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Maturation Medium ◽  
Significant Difference ◽  
Maturation Rate ◽  
Vitro Production

ABSTRACT We aimed to assess the effects of melatonin in the in vitro production of bovine embryos. Our experiment was conducted at the Laboratório de Reprodução Animal of the Universidade Estadual do Maranhão. The cumulus-oocyte complexes (COCs) were distributed among treatments at concentrations of 0, 10-1, 10-3 and 10-5 µMol/L melatonin. Our experiment was further divided into two: the first was to assess the effect of different concentrations of melatonin (treatments) on the maturation rate of COCs, and the second was to assess the effects of melatonin treatments on the in vitro production of bovine embryos. The results from the first experiment demonstrated no significant difference between the in vitro maturation rate of the cultivated COCs in treatments with melatonin. In the second experiment, however, melatonin treatments yielded statistically higher cleavage, morula and blastocyst rates in the 10-5 µM group (52.9%, 52.9%, and 35.3%, respectively), and lower rates in the 10-1 µM group (19.5%, 19.5% and 7.8%, respectively), compared to the others. The control group (no melatonin) and the 10-3 µM group showed similar results. We concluded that supplementation of melatonin in the in vitro maturation medium resulted in no improvement in the oocyte maturation rate, but in the in vitro production of embryos at different concentrations, the 10-5 µM group displayed better results, but with no improvement in the variables (P < 0.05).

scholarly journals A 38 h interval between hCG priming and oocyte retrieval increases in vivo and in vitro oocyte maturation rate in programmed IVM cycles

2008 ◽  
Vol 23 (9) ◽  
pp. 2010-2016 ◽  
Author(s):  
W.-Y. Son ◽  
J.-T. Chung ◽  
R.-C. Chian ◽  
B. Herrero ◽  
E. Demirtas ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Oocyte Retrieval ◽  
Maturation Rate ◽  
In Vitro Oocyte Maturation

scholarly journals THE EFFECT OF UREA SUPLEMENTATION IN MATURATION MEDIA OF BOVINE OOCYTE IN VITRO TOWARDS EXPRESSION OF BAX, BCL-2 AND BAX/BCL-2 RATIO

2020 ◽  
Vol 14 (1) ◽  
Author(s):  
Dhesy Kartikasari ◽  
Sri Mulyati ◽  
Suzanita Utama ◽  
Pudji Srianto ◽  
Widjiati Widjiati ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Immunocytochemical Staining ◽  
Control Group ◽  
Bovine Oocyte ◽  
Significant Difference ◽  
In Vitro Oocyte Maturation ◽  
Positive Results

This study aims to evaluate the expression of BAX, BCL-2, and BAX/BCL-2 ratio in maturation media of cow oocytes which supplemented with Urea in vitro because BAX and BCL-2 are the main regulators of apoptosis. A total of 263 oocytes from follicle aspirations originating from ovaries taken from slaughterhouses and were saturated with 3 addition of urea which was divided into three groups. The control group (P0) was control group without the addition of urea, P1 group was added with urea 20 mg/dL, while P2 group was added with urea 40 mg/dL. The results of in vitro oocyte maturation were continued with identification using immunocytochemical staining with the addition of BAX and BCL-2 antibodies. Positive results showed a brownish color on the oocyte and its cumulus. The results of this study indicated that there were significant differences (P0.05) in BAX and BCL-2 expression, although both curves were equally increase. The increase in BCL-2 was more significant than BAX, while the BAX BCL-2 ratio did not show a significant difference (P0.05) in whichthe curve of BAX/BCL-2 ratio was decreased. It can be concluded that the addition of urea does not affect the level of apoptosis.

187 9-cis RETINOIC ACID IMPROVES MATURATION RATE AND ALTERS GENE EXPRESSION OF IN VITRO-MATURED OOCYTES IN EGYPTIAN BUFFALO

2017 ◽  
Vol 29 (1) ◽  
pp. 202
Author(s):  
A. Gad ◽  
S. Abu Hamed ◽  
M. Khalifa ◽  
A. El-Sayed ◽  
S. A. Swiefy ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Retinoic Acid ◽  
Vitamin A ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Control Group ◽  
Maturation Rate

Retinoic acid, a metabolite of vitamin A, regulates oocyte maturation through multiple mechanisms, including gene expression modulation or preventing oxidative stress. Effects of retinoic acid during oocyte maturation have been reported in several species; however, there have been no studies illustrating these effects in buffalo. Therefore, the objective of this study was to investigate the influence of 9-cis retinoic acid (9-cisRA), an active metabolite of vitamin A, on maturation rate and gene expression during in vitro maturation of buffalo oocytes. Cumulus-oocyte complexes (n = 360) were aspirated from surface follicles of Buffalo ovaries collected from local abattoirs and transported to the laboratory in physiological saline (0.9% NaCl) containing antibiotics (100 µg mL−1 of streptomycin sulfate and 100 IU mL−1 of penicillin) and maintained at 30°C. Grade A cumulus-oocyte complexes (evenly granulated cytoplasm and surrounded by multiple layers of cumulus cells) were randomly divided into 4 groups (90 oocytes/group) and allocated in TCM-199 medium supplemented with 10% fetal bovine serum, 0.2 mM sodium pyruvate, 50 μg mL−1 of gentamycin, and 10 μg mL−1 of FSH and contained 0 (control), 5, 50, or 200 nM of 9-cisRA for maturation. After 24 h, maturation rate was calculated as a percentage based on polar body extrusion. In addition, gene expression patterns were analysed for antioxidant related genes (SOD1, CAT, GPX4, HOMX1, and PRDX1) and oocyte quality-related genes (GDF9 and BMP15) using quantitative real-time PCR with GAPDH as a housekeeping gene. Fold changes (FC) were calculated using ΔΔCt method (FC ≥2; P < 0.05). The results showed that maturation rate (based on the extrusion of polar body) was significantly higher in 5 nM 9-cisRA oocyte group (49.4 ± 2.1%) compared with the control group (35 ± 1.8%); in contrast, the 200 nM 9-cisRA oocyte group showed the lowest maturation rate (27.2 ± 2.7%). However, the 50 nM 9-cisRA oocyte group showed no significant differences (31.2 ± 3.8%) compared with control group .Oocytes treated with 5 and 50 nM 9-cisRA during in vitro maturation showed significant up-regulation of SOD1 (3.4 and 3.08 FC), CAT (2.7 and 1.8 FC), and HOMX1 (4.5 and 4 FC), and significant down-regulation of BMP15 (−3.7 and −3.6 FC), respectively, compared with the control group. Moreover, GPX4, PRDX1, and GDF9 genes were highly expressed in the 50 nM compared with the control group (13.2, 10.4, and 1.8 FC, respectively). In contrast, the 200 nM 9-cisRA group showed significant down-regulation of CAT (−60.3 FC), GDF9 (−2.5 FC), and BMP15 (−9.7 FC) compared with the control group. In conclusion, these results suggested that a low concentration of 9-cisRA (5 nM) in maturation media can improves maturation rate of buffalo oocytes and up-regulates the expression of oxidative stress response-related genes.

scholarly journals Exposure to cell phone induce oxidative stress in mice preantral follicles during in vitro cultivation: An experimental study

2019 ◽  
Author(s):  
Najmeh Vafere Koohestani ◽  
Saeed Zavareh ◽  
Taghi Lashkarbolouki ◽  
Fariba Azimipour
Keyword(s):  
Oxidative Stress ◽  
Cell Phone ◽  
Ovarian Follicle ◽  
Developmental Competence ◽  
Control Group ◽  
In Vitro Cultivation ◽  
Enzymatic Antioxidants ◽  
Preantral Follicles ◽  
Background Radiations

Background: Radiations emitting from mobile phones have been proposed to affect people’s health, mediated by various mechanisms like induction of oxidative stress. Objective: This study aims to investigate the effect of cell phone exposure on the oxidative status of mice preantral follicles (PFs) during in vitro culture. Materials and Methods: PFs (n = 2580) were isolated mechanically from 16 to 18 day-old NMRI mice (n = 50) and divided into control and cell phone-exposed groups. PFs were cultured for 12 days and ovulation was induced using human chorion gonadotropin. The developmental parameters including size, survival, antral cavity formation, ovulation and oocyte maturation were assessed. In parallel, enzymatic antioxidants activities, total antioxidant capacity (TAC), and Malondialdehyde (MDA) levels were evaluated. Results: The diameters and the rates of survival, antrum formation, ovulation, and metaphase II oocytes of exposed PFs to cell phone were significantly lower than those of the control group (p ≤0.001). The PFs exposed to cell phone had significantly lower superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) activity compared with the control group. In the cell phone exposed PFs, the TAC level was significantly lower (p≤ 0.001) and MDA levels were significantly higher (p≤ 0.001), compared to the those of control group. Conclusion: Exposure to cell phone compromised the developmental competence of mice PFs by increasing oxidative stress. Key words: Bovine, Embryonic development, Niacin, Oocytes, VitrificationKey words: Ovarian follicle, Cell phone, Oxidative stress, Mice.

In vitro maturation of oocytes from non-stimulated common wombats

2003 ◽  
Vol 15 (5) ◽  
pp. 303 ◽  
Author(s):  
M. Cleary ◽  
M. West ◽  
J. Shaw ◽  
G. Jenkin ◽  
A. Trounson
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Cumulus Cells ◽  
In Vitro Fertilisation ◽  
Oocyte Collection ◽  
Common Wombat ◽  
Maturation Rate ◽  
Reproductive Techniques ◽  
The Common

Assisted reproductive techniques, such as in vitro oocyte maturation in conjunction with in vitro fertilisation, may be used as a tool to manipulate reproduction. Using the common wombat as a model for the critically endangered northern hairy-nosed wombat, the present study examined whether oocyte maturation could be achieved under field conditions. At the time of collection, no oocytes were at the metaphase II (MII) stage (0/42). After 60 h culture using the submarine incubation system, 34% of oocytes (24/70) matured to telophase/MII, as indicated by the presence of a polar body. The proportion of oocytes that reached MII was higher for oocytes collected from follicles >2 mm in diameter compared with follicles <2 mm (40% v. 22%, respectively). The presence of cumulus cells alone did not influence the maturation potential. Oocytes without cumulus cells collected from follicles >2 mm in diameter had the highest maturation rate (58%). Maturation was not affected by the reproductive status of the common wombat or a delay of up to 5 h before oocyte collection. In conclusion, the present study demonstrated that oocytes collected from non-stimulated common wombats can mature to MII in culture.

scholarly journals Toxicity effect of Auxemma oncocalyx fraction and its active principle oncocalyxone A on in vitro culture of caprine secondary follicles and in vitro oocyte maturation

2017 ◽  
Vol 38 (3) ◽  
pp. 1361 ◽  
Author(s):  
Johanna Leiva-Revilla ◽  
Jesús Cadenas ◽  
Luis Alberto Vieira ◽  
Claudio Cabral Campello ◽  
Juliana Jales de Hollanda Celestino ◽  
...  
Keyword(s):  
Growth Rate ◽  
In Vitro Culture ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cellular Proliferation ◽  
Control Group ◽  
Antral Follicles ◽  
Chromatin Configuration ◽  
In Vitro Oocyte Maturation

Crude extract of the heartwood of Auxemma oncocalyx (A. oncocalyx) and its main component i.e., Oncocalyxone A (onco A), have elevated antioxidant and anti-tumoral activity, but studies on the action of these drugs regarding folliculogenesis are lacking. The aim of this study was to evaluate the effect of A. oncocalyx and onco A on the in vitro culture of isolated secondary follicles and on the in vitro maturation of oocytes from caprine antral follicles grown in vivo. Isolated secondary follicles were randomly distributed in six groups; the non-cultured control was immediately fixed upon isolation. The remaining follicles were cultured for 7 days in ?-MEM+ alone (control) or supplemented with DMSO, doxorrubicin, A. oncocalyx or onco A. After culture, follicles were evaluated for antrum formation, growth rate, apoptosis (TUNEL) and cellular proliferation (PCNA), as well as gene expression of Bcl2 and Bax. Additionally, cumulus oocyte complexes (COCs) were aspirated and allocated into five treatments for in vitro maturation: control, cultured only in maturation base medium (TCM 199+); or supplemented with DMSO; DXR; A. oncocalyx or onco A. After in vitro maturation, oocyte chromatin configuration and viability were assessed. After 7 days of culture, there was a reduction (P < 0.05) in the percentage of morphologically intact follicles, antrum formation, growth rate and number of PCNA positive granulosa cells in DXR treatment compared to the other treatments. In the DXR treatment a higher percentage (P < 0.05) of TUNEL positive follicles and higher (P < 0.05) relative BAX:BCL2 mRNA ratio’s were observed. After in vitro maturation of the COCs DXR, A. oncocalyx and onco A treatments had a greater (P < 0.05) percentage of abnormal oocytes and a lower (P < 0.05) percentage of viable oocytes as compared with the control group. However, only DXR and onco A treatments increased (P < 0.05) the percentage of alive oocytes with abnormal chromatin configuration. There were no differences in maturation rates between the control group and DXR, A. oncocalyx and onco A treatments. In conclusion, under our culture conditions, A. oncocalyx and onco A do not exhibit a toxic effect on isolated secondary follicles and on maturation rates of COCs recovered from antral follicles, however, these drugs negatively affected the COCs viability.  Thus, the use of culture biotechnologies as an in vitro secondary follicle culture and in vitro oocyte maturation toxicity testing are appropriated methods to evaluate the possible effects of drugs in folliculogesis.

scholarly journals 46 Effect of the addition of α-tocopherol to in vitro maturation media of bovine oocytes

2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 2-3
Author(s):  
Theisy P Acosta Pérez
Keyword(s):  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Control Group ◽  
Chi Square ◽  
Cumulus Expansion ◽  
Minimum Concentration ◽  
Maturation Rate ◽  
Chi Square Test ◽  
System Data

Abstract α-tocopherol is known to be a powerful antioxidant, in this regard, it was added to bovine oocyte in vitro maturation media to evaluate its effect on oocyte maturation. Oocytes (n = 624) aspirated from ovaries of slaughtered cows were classified by quality and divided in four categories according to cytoplasm appearance and cumulus cells layers. Oocytes were washed in TCM-199 supplemented with fetal bovine serum (FBS) and FSH, then distributed in maturation media (TCM-199 supplemented with FBS, FSH and gentamicin). Three experimental groups of α-tocopherol (50, 100 and 200 mM) and a control group without α-tocopherol were used. Maturation was carried 22 h at 38.5°C in a 5% CO2 atmosphere. Oocytes were examined to determine cumulus expansion as categorical data (expansion or no expansion), as well as cumulus expansion Index (CEI). For CEI determination oocytes were graded 0 to 4, being 0 those with null expansion and 4 those with a noticeable cell expansion, then the number of oocytes were multiplied by the grade given and a sum of the totals was obtained, the new total was divided by the total of oocytes in the group and the result obtained corresponded to the CEI of the group. Results were analyzed with Chi Square test (for maturation rates) and an ANOVA (for the CEI) using the SAS system, data are presented as mean ± standard error. There was no statistical difference between control and α-tocopherol groups (P &gt;0.05). Numerically, the control group showed a higher maturation rate (100%) and obtained a higher CEI (2.44±0.20), followed by the 50 mM group (98.16%; 2.39±0.13), the groups 200 mM (97.40%; 2.00±0.14) and 100 mM (96.25%; 2.06±0.24) were the lowest. The addition of the minimum concentration (50 mM) of α-tocopherol to the maturation media could improve maturation rates without exposing oocytes to toxic effects.

Supplementation of c-type natriuretic peptide during in vitro growth period benefits the development of murine preantral follicles

Zygote ◽  
2020 ◽  
pp. 1-5
Author(s):  
Li Ang ◽  
Cao Haixia ◽  
Li Hongxia ◽  
Li Ruijiao ◽  
Guo Xingping ◽  
...  
Keyword(s):  
Natriuretic Peptide ◽  
Granulosa Cells ◽  
Culture System ◽  
Early Stage ◽  
Developmental Competence ◽  
Control Group ◽  
Follicle Development ◽  
Control Groups ◽  
Preantral Follicles

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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