Effects of bone morphogenetic protein 4 (BMP4) on in vitro development and survival of bovine preantral follicles enclosed in fragments ovarian tissue

Zygote ◽  
2017 ◽  
Vol 25 (3) ◽  
pp. 256-264 ◽  
Author(s):  
Ellen de Vasconcelos da Cunha ◽  
Glaucinete Borges de Souza ◽  
José Renato de Sousa Passos ◽  
Anderson Weiny Barbalho Silva ◽  
Andressa Minussi Dau ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Culture Medium ◽  
Ovarian Tissue ◽  
Bone Morphogenetic Protein 4 ◽  
Mrna Levels ◽  
Follicle Growth ◽  
In Vitro Development ◽  
Morphogenetic Protein ◽  
Vitro Development

SummaryThe aim of this study was to evaluate the effects of different concentrations of BMP4 on activation, development and mRNA expression of GDF9, BMP15, PCNA, Bax and Bcl2 in cultured bovine follicles enclosed in ovarian tissues. Ovarian tissue fragments were cultured for 6 days in α-MEM+ alone or supplemented with different concentrations of BMP4 (10, 50 or 100 ng/ml). Classical histology was performed to analyze follicle growth and morphology, while real-time PCR was used to analyze mRNA levels in fresh and cultured tissues. After 6 days, the culture of ovarian tissue in α-MEM+ alone or supplemented with 10, 50 or 100 ng/ml BMP4 promoted follicular activation. The different concentrations of BMP4 maintained the percentage of normal follicles similar to results of the control. The presence of 100 ng/ml BMP-4 in culture medium increased oocyte and follicular diameters of primary and secondary follicles when compared with those follicles from uncultured control or cultured in α-MEM+ alone (P < 0.05). The tissues cultured in the presence of increasing concentrations of BMP4 had an increase in mRNA expression of the tested genes, but despite this the differences were not statistically significant. In conclusion, 100 ng/ml BMP4 promotes an increase in diameters of follicles and oocytes of primary and secondary follicles after 6 days of in vitro culture.

In Vitro Development and mRNA Expression in Bovine Nuclear Transferred Embryos Derived from Early G1 or G0 Phase Cells.

2008 ◽  
Vol 78 (Suppl_1) ◽  
pp. 130-131
Author(s):  
Kazutsugu Matsukawa ◽  
Satoshi Akagi ◽  
Kazuhiro Hukunari ◽  
Fukashi Inoue ◽  
Tadashi Furusawa ◽  
...  
Keyword(s):  
Mrna Expression ◽  
In Vitro Development ◽  
G0 Phase ◽  
Vitro Development

Effect of LH and Estradiol-17β Supplementation at Different Time Points on In vitro Development of Preantral Follicles in Sheep

2020 ◽  
Author(s):  
L.S.S. Varaprasad Reddy ◽  
B.R. Naik ◽  
A.V.N. Sivakumar ◽  
B. Punyakumari ◽  
J. Suresh
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Follicular Development ◽  
Standard Medium ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Time Points ◽  
Vitro Development ◽  
Estradiol 17Β

Background: Ovarian follicular development and growth are controlled by many hormones and growth factors. Despite the fact that LH and estradiol-17β have been utilized for the in vitro culture of preantral follicles yet, the suitable time points of supplementation of LH and estradiol-17β is not known. Therefore this study aimed to investigate the influence of addition of LH and estradiol-17β at different time points on in vitro development of preantral follicles (PFs’) in sheep. Method: Preantral follicles isolated from the ovarian cortical slices using micro dissection method were cultured for six days in Bicarbonate buffered Tissue culture medium 199B (TCM 199B) or in a standard culture medium supplemented with LH (2 μg/ml) and estradiol-17β (5 ng/ml) at different points during the culture period. COCs isolated from the follicles at the end of six day culture in different treatments were subjected to in vitro maturation for additional 24h. Result: Supplementation of LH and estradiol-17β during last two days of the culture supported better proportion of PFs’ exhibiting growth whereas supplementation of LH and estradiol-17β during first two days of the culture supported better average increase in diameter and proportion of PFs’ exhibiting antrum formation at the end of six day culture. Further the oocytes in COCs isolated at the end of culture in these treatments and subsequently subjected to in vitro maturation (IVM) for 24hr developed at a higher frequency to MII (metaphase II) stage. Supplementation of LH and estradiol-17β to TCM 199B culture medium in early stages followed by standard medium alone in later stages supports better development of PFs’ in vitro. Following supplementation with LH and estradiol-17β for the first two days culture of PFs’ in standard medium appears to be advantageous for the development of preantral follicles in vitro.

Effects of growth differentiation factor-9 and FSH on in vitro development, viability and mRNA expression in bovine preantral follicles

2013 ◽  
Vol 25 (8) ◽  
pp. 1194 ◽  
Author(s):  
G. L. Vasconcelos ◽  
M. V. A. Saraiva ◽  
J. J. N. Costa ◽  
M. J. Passos ◽  
A. W. B. Silva ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Nuclear Antigen ◽  
Mrna Levels ◽  
Related Factors ◽  
Differentiation Factor ◽  
Growth Differentiation Factor 9 ◽  
Before And After ◽  
Vitro Development

The present study investigated the role of growth differentiation factor (GDF)-9 and FSH, alone or in combination, on the growth, viability and mRNA expression of FSH receptor, proliferating cell nuclear antigen (PCNA) and proteoglycan-related factors (i.e. hyaluronan synthase (HAS) 1, HAS2, versican, perlecan) in bovine secondary follicles before and after in vitro culture. After 12 days culture, sequential FSH (100 ng mL–1 from Days 0 to 6 and 500 ng mL–1 from Days 7 to 12) increased follicular diameter and resulted in increased antrum formation (P < 0.05). Alone, 200 ng mL–1 GDF-9 significantly reduced HAS1 mRNA levels, but increased versican and perlecan mRNA levels in whole follicles, which included the oocyte, theca and granulosa cells. Together, FSH and GDF-9 increased HAS2 and versican (VCAN) mRNA levels, but decreased PCNA mRNA expression, compared with levels in follicles cultured in α-minimum essential medium supplemented with 3.0 mg mL–1 bovine serum albumin, 10 µg mL–1 insulin, 5.5 µg mL–1 transferrin, 5 ng mL–1 selenium, 2 mM glutamine, 2 mM hypoxanthine and 50 μg mL–1 ascorbic acid (α-MEM+). Comparisons of uncultured (0.2 mm) and α-MEM+ cultured follicles revealed that HAS1 mRNA expression was higher, whereas VCAN expression was lower, in cultured follicles (P < 0.05). Expression of HAS1, VCAN and perlecan (HSPG2) was higher in cultured than in vivo-grown (0.3 mm) follicles. In conclusion, FSH and/or GDF-9 promote follicular growth and antrum formation. Moreover, GDF-9 stimulates expression of versican and perlecan and interacts positively with FSH to increase HAS2 expression.

Bovine dominant follicular fluid promotes the in vitro development of goat preantral follicles

2012 ◽  
Vol 24 (3) ◽  
pp. 490 ◽  
Author(s):  
A. B. G. Duarte ◽  
V. R. Araújo ◽  
R. N. Chaves ◽  
G. M. Silva ◽  
D. M. Magalhães-Padilha ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Culture Medium ◽  
Follicular Development ◽  
Ultrastructural Analysis ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Nuclear Membranes ◽  
Vitro Development ◽  
First Time

The aim of this study was to evaluate the effect of follicular fluid collected from bovine dominant follicles (bFF) on the in vitro development of goat preantral follicles and determine the best time to add this supplement to the culture medium. The preantral follicles were isolated and randomly distributed into four treatments in absence (control) or presence of 10% of bFF added on Days 0 (FF0–18), 6 (FF6–18) or 12 (FF12–18) of culture onwards. After 18 days, follicular development was assessed based on follicular survival, antral cavity formation, increased follicular diameter as well as fully grown oocyte (>110 μm) viability and meiosis resumption. The oocytes from the cultured follicles were in vitro-matured and processed for fluorescence or ultrastructural analysis. The results showed that on Day 18 the treatment FF0–18 had a significantly higher (P < 0.05) survival than control and FF12–18, but not FF6–18. The addition of bFF at the beginning of culture (FF0–18 and FF6–18) promoted a high percentage of follicular growth, meiosis resumption and early antrum formation. Moreover, this study described for the first time the ultrastructural analysis of caprine oocytes grown in vitro. This evaluation revealed that in the presence of bFF on (FF0–18) the in vitro-grown oocytes presented normal organelle distribution and well-defined, intact plasma and nuclear membranes. In conclusion, bFF originating from dominant follicles maintain the survival and promote the in vitro growth of goat preantral follicles when added at the beginning of culture.

Sequential medium with GH and IGF-1 improved in vitro development of bovine preantral follicles enclosed in ovarian tissue

2018 ◽  
Vol 53 (5) ◽  
pp. 1103-1113 ◽  
Author(s):  
Carolina Rodriguez Jimenez ◽  
Jovana Luiza de Azevedo ◽  
Torres Ciro Alexandre Alves ◽  
Jurandy Mauro Penitente-Filho ◽  
Wagner Gonzaga Gonçalves
Keyword(s):  
Ovarian Tissue ◽  
In Vitro Development ◽  
Preantral Follicles ◽  
Vitro Development
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