scholarly journals Successful Piglet Production in a Chemically Defined System for In-vitro Production of Porcine Embryos: Dibutyryl Cyclic AMP and Epidermal Growth Factor-family Peptides Support In-vitro Maturation of Oocytes in the Absence of Gonadotropins

2009 ◽  
Vol 55 (4) ◽  
pp. 446-453 ◽  
Author(s):  
Yuka AKAKI ◽  
Koji YOSHIOKA ◽  
Michiko NOGUCHI ◽  
Hiroyoshi HOSHI ◽  
Hiroaki FUNAHASHI
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cyclic Amp ◽  
In Vitro Maturation ◽  
Dibutyryl Cyclic Amp ◽  
In Vitro Production ◽  
Epidermal Growth ◽  
Vitro Production

Epidermal growth factor increases inhibin synthesis by isolated segments of rat seminiferous tubules

1989 ◽  
Vol 123 (2) ◽  
pp. 213-219 ◽  
Author(s):  
G. F. Gonzales ◽  
G. P. Risbridger ◽  
D. M. de Kretser
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cyclic Amp ◽  
Phorbol Esters ◽  
Seminiferous Tubules ◽  
Dibutyryl Cyclic Amp ◽  
Adult Rats ◽  
Adult Rat ◽  
Epidermal Growth

ABSTRACT The effect of epidermal growth factor (EGF) on the production of immunoreactive inhibin by adult rat isolated seminiferous tubules in vitro has been investigated. EGF (0·1–1000 ng/ml) added to cultures of seminiferous tubules from adult rats caused a dose-dependent increase in inhibin content in the tubules without changing the amount secreted into the media. However, after continuous stimulation with EGF for periods in excess of 5 days, an increase in inhibin secretion was observed. In the presence of 10 and 100 ng FSH/ml, EGF (10 ng/ml) produced a further increment in the inhibin content of the tubules, but this effect was not found with FSH concentrations of 500 or 1000 ng/ml. EGF also increased the tubule content of inhibin after the addition of 100 μg dibutyryl cyclic AMP/ml but no effect of EGF was observed on the FSH- or dibutyryl cyclic AMP-induced secretion of inhibin into the medium. The effect of EGF on inhibin content in the tubules was partially suppressed by the addition of 4β-phorbol-12β-myristate-13α-acetate (20 ng/ml). Insulin (1–100 ng/ml) decreased basal inhibin secretion without changing the inhibin content of tubules and this effect was antagonized by EGF (10 ng/ml) with insulin doses of 1–50 ng/ml whereas, at 100 ng/ml, the effect of EGF on tubule inhibin content was reversed. The addition of EDTA (2 mmol/l) resulted in an inhibition of basal and EGF-induced inhibin production. These data demonstrate a stimulatory effect of EGF on inhibin production by isolated seminiferous tubules which is inhibited by insulin and phorbol esters, both stimulators of protein kinase C activity. Journal of Endocrinology (1989) 123, 213–219

79 EFFECTS OF EPIDERMAL GROWTH FACTOR, β-MERCAPTOETHANOL, GLUCOSE, OXYGEN CONCENTRATION, AND FIBROBLAST SUBCULTURE ON THE DEVELOPMENT OF PORCINE NT EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 157
Author(s):  
H. B. Seok ◽  
J. H. Quan ◽  
S. K. Kim
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Developmental Rate ◽  
Blastocyst Stage ◽  
First Polar Body ◽  
Fetal Fibroblasts ◽  
O2 Concentration ◽  
Epidermal Growth

The purpose of this study was to investigate in vitro maturation rate of oocytes cultured in maturation medium supplemented with epidermal growth factor (EGF), β-mercaptoethanol (ME), and glucose, and the further development of NT embryos under various conditions. The basic media used for oocyte maturation were NCSU-23 and PZM-3 supplemented with 0.1 mg mL-1 cysteine, 10% (v/v) porcine follicular fluid (pFF), 10 �g mL-1 FSH, 10 �g mL-1 LH, 20 ng mL-1 EGF, and 25 �M ME. Porcine ovaries were collected at a local slaughterhouse, and donor cells from a 35-day-old fetus were dissociated, resuspended, and cultured for 6–8 days in DMEM supplemented with 10% (v/v) FBS, penicillin G (75 �g mL-1), streptomycin (50 �g mL-1), 1 mM sodium pyruvate, and 1% (v/v) nonessential amino acids. The first polar body and adjacent cytoplasm were enucleated by a micropipette in HEPES-buffered NCSU-23 supplemented with 4 mg mL-1 BSA and 7.5 �g mL-1 cytochalasin B. Couplets were equilibrated with 0.3 M mannitol solution and transferred to a chamber containing 2 electrodes with a pulse of 2.1 kV cm-1 for 30 �s. When the embryos were cultured in NCSU-23 or PZM-3 supplemented with or without 20 ng mL-1 EGF for 144 h, the development rates to the blastocyst stage were 12.0 � 1.3%, 9.6 � 1.9%, 10.9 � 2.1%, and 9.1 � 2.3%, respectively. When the embryos were cultured in NCSU-23 or PZM-3 supplemented with or without 25 �M ME for 144 h, the rates to blastocyst stage were 9.6 � 1.7%, 7.3 � 2.3%, 11.9 � 1.8%, and 7.4 � 2.1%, respectively. The developmental rate to the blastocyst stage of NT embryos cultured in PZM-3 supplemented with ME was significantly higher than when cultured without ME supplementation (P < 0.05). When the embryos were cultured in NCSU-23 or PZM-3 supplemented with or without 1.5 mM glucose for 144 h, the rates to blastocyst stage were 9.4 � 2.2%, 6.8 � 2.7%, 10.9 � 2.4%, and 8.9 � 2.6%, respectively. The developmental rate to the blastocyst stage of NT embryos cultured in NCSU-23 and PZM-3 supplemented with glucose was higher than when cultured without glucose supplementation. When NT embryos were cultured in NUSU-23 and PZM-3 at 5% and 20% O2 concentration, the rates were 11.1 � 1.8%, 9.8 � 1.4%, 12.5 � 1.6%, and 10.9 � 1.5%, respectively. The developmental rate to the blastocyst stage of NT embryos cultured in both NCSU-23 and PZM-3 at 5% O2 concentration was higher than when cultured at 20% O2 concentration. When fetal fibroblasts were cultured in NCSU-23 and PZM-3, the fusion rate of less than 10 passages was higher than for those of 11–15 passages. In conclusion, the present study indicates that EGF and glucose have beneficial effects on the in vitro maturation of oocytes, and ME improves the developmental ability of NT embryos. Furthermore, the developmental rate in subcultured fibroblast cells was improved when reconstruction was made with less than 10 passages.

scholarly journals Comparison of rapid changes in surface morphology and coated pit formation of PC12 cells in response to nerve growth factor, epidermal growth factor, and dibutyryl cyclic AMP.

1984 ◽  
Vol 98 (2) ◽  
pp. 457-465 ◽  
Author(s):  
J L Connolly ◽  
S A Green ◽  
L A Greene
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cyclic Amp ◽  
Cell Surface ◽  
Dibutyryl Cyclic Amp ◽  
Coated Pits ◽  
Pit Formation ◽  
Epidermal Growth ◽  
Surface Changes

Scanning and transmission electron microscopic studies were carried out on the rapid cell surface response of PC12 pheochromocytoma cells to treatment with nerve growth factor (NGF), epidermal growth factor (EGF), and dibutyryl cyclic AMP. EGF induced a rapidly initiated series of surface changes identical to those previously observed with NGF. Ruffles appear over the dorsal surface of the cells by 30 s, are prominent at 3 min, and are absent by 7 min. Microvilli disappear as dorsal ruffles become prominent. Peripheral ruffles are seen by 3 min, are prominent on most of the cells by 7 min, and are virtually absent by 15 min. Large blebs are present on 50% of the cells by 2 h and are markedly decreased by 4 h. Within 30 s after NGF or EGF addition, an increase in the density of 60-130-nm coated pits per unit membrane is detectable. This reaches a maximum of two- to threefold in from 1 to 3 min and gradually decreases. Combined treatment with NGF and EGF increases surface ruffling and, after an early peak in coated pits which at 3 min is similar in magnitude to that observed for the separately administered factors, maintains a greater number of pits per unit area than either treatment alone. 3-d pretreatment with NGF greatly reduces the response of the cells to EGF both with respect to surface ruffling and coated pit formation while 4-h NGF pretreatment has no effect on the EGF response. Dibutyryl cyclic AMP induced none of the rapidly onsetting changes caused by NGF or EGF, and therefore it seems unlikely that cyclic AMP mediates these surface changes. Changes in cell surface architecture induced by NGF and EGF on PC12 cells and by NGF in normal sympathetic neurons (as previously described) indicates that such responses may be a widespread phenomenon associated with the interaction of at least some peptide growth factors/hormones with their receptors. These responses may represent or reflect primary events in the mechanism by which these factors act.

Sign in / Sign up

Export Citation Format

Share Document