Role of follicle stimulating hormone and epidermal growth factor in the development of porcine preantral follicle in vitro

Zygote ◽  
2007 ◽  
Vol 15 (3) ◽  
pp. 233-240 ◽  
Author(s):  
Ji Wu ◽  
Qi Tian
Keyword(s):  
Growth Factor ◽  
Embryonic Development ◽  
Follicle Stimulating Hormone ◽  
Preantral Follicle ◽  
Preantral Follicles ◽  
Antral Follicles ◽  
Estradiol Secretion ◽  
Epidermal Growth

SummaryThe aim of the present study was to assess the role of follicle stimulating hormone (FSH), epidermal growth factor (EGF) or a combination of EGF and FSH on the in vitro growth of porcine preantral follicles, estradiol secretion, antrum formation, oocyte maturation and subsequent embryonic development. Porcine preantral follicles were cultured for 3 days in the absence or in the presence of FSH or EGF. Oocytes from these follicles were then matured, fertilized in vitro and embryos were cultured. Estradiol secretion and histological analysis of cultured follicles were also carried out. The results showed that when FSH, or a combination of EGF and FSH, was added to the culture medium, most of preantral follicles grew to antral follicles with high estradiol secretion and the oocytes from these antral follicles could mature, fertilize and develop to the blastocyst stage. Without FSH, or a combination of EGF and FSH, preantral follicles were unable to develop to the antral stage. Histology demonstrated that the resulting follicles were nonantral, estradiol production was reduced and none of their oocytes matured after in vitro maturation. The results indicate the essential role of FSH in promoting in vitro growth of porcine preantral follicle, estradiol secretion, antrum formation, oocyte maturation and subsequent embryonic development. EGF with FSH treatment of porcine preantral follicles improves the quality of oocytes, shown by a higher frequency of embryonic development.

Co-culture of buffalo (Bubalus bubalis) preantral follicles with antral follicles: a comparative study of developmental competence of oocytes derived from in vivo developed and in vitro cultured antral follicles

Zygote ◽  
2012 ◽  
Vol 21 (3) ◽  
pp. 286-294 ◽  
Author(s):  
G. Taru Sharma ◽  
Pawan K. Dubey ◽  
Amar Nath ◽  
G. Saikumar
Keyword(s):  
Growth Factor ◽  
Bubalus Bubalis ◽  
Developmental Competence ◽  
Cleavage Rate ◽  
Term Survival ◽  
Preantral Follicles ◽  
Antral Follicles ◽  
Epidermal Growth

SummaryThe present study was undertaken to examine whether the presence of antral follicles (AFs) affects the survival, growth and steroidogenesis of preantral follicles (PFs) and compare the maturation and developmental competence of buffalo oocytes derived from in vivo developed and in vitro cultured AFs. Two experiments were carried out. In experiment I, PFs (200–250 μm) were isolated and cultured with or without AFs (3–5 mm) in TCM-199 medium that contained 10% fetal bovine serum (FBS), 1% insulin transferin selenium (ITS), 20 ng/ml epidermal growth factor (EGF), 0.5 μg/ml follicle-stimulating hormone (FSH) and 100 ng/ml insulin-like growth factor (IGF)-I. In experiment II, in vitro developmental competence was compared for the cumulus–oocyte complexes (COCs) recovered from in vivo developed and in vitro cultured AFs. Survival, growth, development of antrum, accumulation of estradiol and progesterone was (P < 0.05) higher when PFs were co-cultured with AFs. Developmental competence of both types of follicular oocytes did not differ significantly in terms of maturation and cleavage rate, but morula and blastocyst production rate were (P < 0.05) higher with in vivo developed AFs as compared with the in vitro cultured antral follicular oocytes. In conclusion, co-culture of PFs with AFs supports long-term survival and growth of buffalo PFs and this co-culture system plays a dual role for in vitro production of embryos as well as understanding the relationship between developing PFs and AFs.

In vitro culture of oocytes with surrounding cumulus complexes and granulosa cells (COCGs) from bovine early antral follicles

2003 ◽  
Vol 77 (1) ◽  
pp. 141-147
Author(s):  
S. Saha ◽  
M. Shimizu ◽  
M. Geshi ◽  
Y. Izaike
Keyword(s):  
Growth Factor ◽  
Granulosa Cells ◽  
Culture Medium ◽  
Follicle Stimulating Hormone ◽  
Germinal Vesicle ◽  
Antral Follicles ◽  
Ovarian Cortex ◽  
Epidermal Growth ◽  
Germinal Vesicle Break Down

AbstractCumulus-oocyte complexes with surrounding granulosa cells (COCGs) in early antral follicles (0·5 to 0·7 mm in diameter) were surgically collected from sections of bovine ovarian cortex under a dissection microscope and subsequently cultured in vitro using follicle stimulating hormone (FSH), epidermal growth factor (EGF), insulin-transferrin-selenium (ITS) and hypoxanthine, singly or in combination, to obtain fully grown matured oocytes. Oocytes cultured in the presence of FSH + hypoxanthine increased (P < 0·05) in diameter from 93 µm on the day of commencement of culture to 106·37±0·34 µm on day 5. Oocytes cultured in the presence of FSH, hypoxanthine or hypoxanthine + ITS + FSH increased (P < 0·05) to mean diameters of 105·40 (s.e. 0·47) µm, 105·50 (s.e. 0·39) µm and 105·35 (s.e. 0·55) µm, respectively. By day 11 of culture, oocyte diameters 110·50 (s.e. 0·35) µm, 110·13 (s.e. 0·39) µm, 109·49 (s.e. 0·46) µm, 109·53 (s.e. 0·58) µm and 109·16 (s.e. 0·43) µm were recorded for treatments FSH + hypoxanthine, hypoxanthine + ITS + FSH, FSH, hypoxanthine and FSH + EGF + hypoxanthine + ITS, respectively. The proportions with COCGs which formed an antrum while cultured in vitro; were categorized as morphologically normal following recovery from the gel; matured in vitro; showed germinal vesicle break down and reached metaphase II were highest (P < 0·05) for the FSH + hypoxanthine treatment (49/60 (81·7%), 48/60 (80·0%), 47/60 (78·3%), 45/60 (75·0%) and 15/60 (25·0%), respectively, followed by hypoxanthine + ITS + FSH (47/60 (78·3%), 44/60 (73·3%), 41/60 (68·3%), 41/60 (68·3%) and 12/60 (20%), respectively), FSH (43/60 (71·7%), 42/60 (70%), 40/60 (66·7%), 39/60 (65·0%) and 9/60 (15%), respectively) and hypoxanthine (41/60 (68·3%), 38/60 (63·3%), 36/60 (60%), 35/60 (58·3%) and 8/60 (13·3%), respectively). In experiment II, the in vitro fertilization and cleavage rates of COCGs were highest (P < 0·05) for FSH + hypoxanthine treatment (17/60; 28·3%) followed by hypoxanthine + ITS + FSH (13/60; 21·6%), FSH (12/60; 20%) and hypoxanthine (11/60; 18·3%) treatments. The results of this study show that COCGs from early antral follicles can be isolated, cultured and grown in vitro. Furthermore, supplements like FSH and hypoxanthine can be used singly or in combination(s) in culture medium to enhance the growth of COCGs.

Follicle-stimulating hormone-dependent estrogen secretion by rat Sertoli cells in vitro: Modulation by calcium

1991 ◽  
Vol 125 (3) ◽  
pp. 280-285 ◽  
Author(s):  
J. Alan Talbot ◽  
Ann Lambert ◽  
Robert Mitchell ◽  
Marek Grabinski ◽  
David C. Anderson ◽  
...  
Keyword(s):  
Sertoli Cells ◽  
Culture Medium ◽  
Follicle Stimulating Hormone ◽  
Dibutyryl Camp ◽  
Immature Rat ◽  
Estradiol Secretion ◽  
Old Rats ◽  
Stimulating Hormone

Abstract We have investigated the role of Ca2+ in the control of FSH-induced estradiol secretion by Sertoli cells isolated from 8-10 days old rats. Exogenous Ca2+ (4-8 mmol/1) inhibited FSH-stimulated E2 secretion such that, with 8 mmol/l Ca2+ and FSH (8 IU/l) E2 secretion decreased from 2091±322 to 1480±84 pmol/l (p<0.002), whilst chelation of Ca2+ in the culture medium with EGTA (3 mmol/l) increased E2 secretion from 360±45 to 1242±133 pmol/l) in the absence of FSH. Further, EGTA (3 mmol/l) markedly potentiated FSH (8 IU/l), forskolin (1 μmol/l) and dibutyryl cAMP (1 mmol/l)-stimulated E2 secretion. Addition of the Ca2+ ionophores, ionomycin (2-5 μmol/l) and A23187 (2 μmol/l), inhibited FSH (8 IU/l)-stimulated E2 secretion by >80%. The effect of ionomycin was totally reversible, whereas that of A23187 was irreversible. Ionomycin (5 μmol/l) had no effect on EGTA-induced E2 secretion in the absence of FSH, but reduced EGTA-provoked E2 secretion by 59% in the presence of FSH (8 IU/l). Similarly, forskolin- and dibutyryl cAMP-provoked E2 production was inhibited 46-50% by ionomycin (5 μmol/l). We conclude that FSH-induced E2 secretion from immature rat Sertoli cells is modulated by intra- and extracellular Ca2+.

Synergetic effects of epidermal growth factor and estradiol on cytoplasmic maturation of porcine oocytes

Zygote ◽  
2002 ◽  
Vol 10 (4) ◽  
pp. 349-354 ◽  
Author(s):  
Yong-Hai Li ◽  
Rui-Hua Liu ◽  
Li-Hong Jiao ◽  
Wei-Hua Wang
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Early Embryo ◽  
Antral Follicles ◽  
Nuclear Maturation ◽  
17Β Estradiol ◽  
Epidermal Growth ◽  
Cytoplasmic Maturation ◽  
Pronuclear Formation

This study was conducted to examine the effect of epidermal growth factor (EGF) and 17β-estradiol (E2) on nuclear and cytoplasmic (male pronuclear formation and early embryo development) maturation of porcine oocytes. Oocytes were aspirated from antral follicles and cultured in modified TCM-199 medium supplemented with 0.57 mM cysteine, 10 IU/ml eCG, 10 IU/ml hCG, with or without EGF and/or E2. In vitro fertilisation of matured oocytes was performed in a modified Tris-buffered medium (mTBM) with frozen-thawed ejaculated spermatozoa. Oocytes were transferred to NCSU-23 supplemented with 0.4% bovine serum albumin at 6 h after in vitro fertilisation. Significantly higher (p < 0.05) rates of nuclear maturation, pronuclear formation and cleavage (91.7%, 65.2% and 37.3%, respectively) were observed when oocytes were cultured in the medium containing both EGF (10 ng/ml) and E2 (1 μg/ml) than in the medium supplemented with either EGF or E2 or without both. Intracellular glutathione concentration in the oocytes cultured in the medium containing both E2 and EGF was also significantly higher (12.1 pmol per oocyte) than that of oocytes cultured in the medium with E2 or EGF alone or without both. These findings suggested that EGF and E2 have a synergestic effect on both nuclear and cytoplasmic maturation of porcine oocytes.

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