scholarly journals Epidermal growth factor effects on marmoset monkey (Callithrix jacchus) oocyte in vitro maturation, IVF and embryo development are altered by gonadotrophin concentration during oocyte maturation

2010 ◽  
Vol 25 (8) ◽  
pp. 2047-2058 ◽  
Author(s):  
O. Y. Tkachenko ◽  
S. Delimitreva ◽  
E. Isachenko ◽  
R. R. Valle ◽  
H. W. Michelmann ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Callithrix Jacchus ◽  
Marmoset Monkey ◽  
Epidermal Growth

scholarly journals Effect of epidermal growth factor-like peptides on pig cumulus cell expansion, oocyte maturation, and acquisition of developmental competence in vitro: comparison with gonadotropins

Reproduction ◽  
2011 ◽  
Vol 141 (4) ◽  
pp. 425-435 ◽  
Author(s):  
Radek Procházka ◽  
Michal Petlach ◽  
Eva Nagyová ◽  
Lucie Němcová
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Oocyte Maturation ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Blastocyst Stage ◽  
Human Chorionic Gonadotrophin ◽  
Developmental Competence ◽  
Epidermal Growth

The aim of this work was to assess the FSH-stimulated expression of epidermal growth factor (EGF)-like peptides in cultured cumulus–oocyte complexes (COCs) and to find out the effect of the peptides on cumulus expansion, oocyte maturation, and acquisition of developmental competencein vitro. FSH promptly stimulated expression of amphiregulin (AREG) and epiregulin (EREG), but not betacellulin (BTC) in the cultured COCs. Expression ofAREGandEREGreached maximum at 2 or 4 h after FSH addition respectively. FSH also significantly stimulated expression of expansion-related genes (PTGS2,TNFAIP6, andHAS2) in the COCs at 4 and 8 h of culture, with a significant decrease at 20 h of culture. Both AREG and EREG also increased expression of the expansion-related genes; however, the relative abundance of mRNA for each gene was much lower than in the FSH-stimulated COCs. In contrast to FSH, AREG and EREG neither stimulated expression ofCYP11A1in the COCs nor an increase in progesterone production by cumulus cells. AREG and EREG stimulated maturation of oocytes and expansion of cumulus cells, although the percentage of oocytes that had reached metaphase II was significantly lower when compared to FSH-induced maturation. Nevertheless, significantly more oocytes stimulated with AREG and/or EREG developed to blastocyst stage after parthenogenetic activation when compared to oocytes stimulated with FSH alone or combinations of FSH/LH or pregnant mares serum gonadotrophin/human chorionic gonadotrophin. We conclude that EGF-like peptides do not mimic all effects of FSH on the cultured COCs; nevertheless, they yield oocytes with superior developmental competence.

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 Effects of oocyte collection techniques and maturation media on in vitro maturation and subsequent embryo development in Boer goat

2008 ◽  
Vol 52 (No. 1) ◽  
pp. 21-25 ◽  
Author(s):  
Z.G. Wang ◽  
Z.R. Xu ◽  
S.D. Yu
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Embryo Development ◽  
The Other ◽  
Boer Goat ◽  
Oocyte Recovery ◽  
Oocyte Collection ◽  
Epidermal Growth ◽  
Alternative Techniques

The oocytes (experiment 1) were harvested by one of the four collection techniques (slicing, puncture, aspiration I and aspiration II) and the total number and the number of each grade of oocytes were counted, respectively. The good-quality oocytes (good and fair grade) were cultured for maturation. In experiment 2, the oocytes were matured in TCM-199 supplemented with 10 ng/ml of epidermal growth factor (EGF) or 10% FCS, either alone or with 1 IU/ml FSH, or the oocytes were matured in TCM-199 supplemented with 1 IU/ml FSH. After maturation, the oocytes in the two experiments were fertilized, respectively. Slicing (6.3) and puncture (5.8) of the ovaries yielded a higher (<i>P</i> < 0.05) number of oocytes per ovary compared to aspiration I (2.9) and aspiration II (3.1). Oocytes matured in the TCM-199 medium supplemented with EGF or FCS with FSH had a significantly higher proportion of blastocysts than the other treatments (<i>P</i> < 0.05). In conclusion, slicing and puncture are alternative techniques of oocyte recovery in Boer goat. The TCM-199 medium supplemented with EGF or FCS in the presence of FSH is suitable for <i>in vitro</i> maturation of oocytes.

343 EFFECTS OF EPIDERMAL GROWTH FACTOR SUPPLEMENTATION ON IN VITRO MATURATION AND GENE EXPRESSION OF CANINE OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 279
Author(s):  
H.-J. Song ◽  
S.-H. Lee ◽  
G.-H. Maeng ◽  
J.-G. Kim ◽  
S. Balasubramanian ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Luteinizing Hormone Receptor ◽  
Control Group ◽  
Dependent Manner ◽  
Junction Protein ◽  
Epidermal Growth

Despite many efforts to improve canine in vitro maturation (IVM), the efficiency is still low compared to that of other mammalian species (Marie et al. 2004). Epidermal growth factor (EGF) has stimulatory effects on the resumption of oocyte maturation and cumulus expansion in vitro and on prei-mplantation embryonic development in mammals by either an autocrine or a paracrine pathway, or a combination of both systems (Paria et al. 2001 PNAS 98, 1047-1052). The present study investigated the effects of EGF supplementation on in vitro maturation and gene expression of canine oocytes. Oocytes were recovered by slicing ovaries recovered from 40 bitches after ovariohysterectomy at random stages of the estrous cycle. Cumulus-oocyte complexes (COCs) were matured in TCM-199 containing 10% FBS, 1 �g/mL FSH and LH, and EGF (0, 10, or 30 ng/mL) for 48 or 72 h at 39�C in a humidified atmosphere of 5% CO2 in air. In Experiment I (n = 2520 oocytes), the nuclear maturation status was assessed by fluorescence microscopy after bisbenzimide (Hoechst 33342) staining (10 �g/mL) at 0, 48, and 72 h of incubation. In Experiment II (n = 90 oocytes), expression of transcripts such as EGF receptor (EGFR), luteinizing hormone receptor (LHR), and gap junction protein (GJA5) were determined in 10 intact COCs each at 0, 48, and 72 h, respectively, by reverse transcription-polymerase chain reaction (RT-PCR). At 0 h 10-20% of the oocytes had undergone resumption of meiosis (GVBD<MII). After 48 h of IVM, rate of meiotic resumption for 0, 10, and 30 ng/mL EGF were 28, 35, and 30%, respectively. At 72 h of IVM, oocytes in the 10 ng/mL EGF group had resumed meiosis at a higher frequency (55%; P < 0.05) than in the 30 ng/mL EGF or the control group (39 and 42%, respectively). At 72 h of IVM, the frequency of maturation to the MII stage was significantly higher in the 10 ng/mL EGF group (9.6%) than in the 30 ng/mL EGF or the control group (4.2 and 3.3%, respectively). The expression of EGFR was significantly higher (P < 0.05) in 0 h oocytes than in the 48- or 72-h oocytes. Further EGFR expression levels were decreased in the presence of EGF in a dose dependent manner. Transcripts for LHR were detected at all maturation intervals and its expression patterns were not altered by supplementation with 10 ng/mL EGF. Expression of GJA5 was observed only after 48 h of IVM, and levels of expression were similar in oocytes supplemented with both 10 and 30 ng/mL EGF. In summary, our results indicate that supplementation of canine IVM medium with 10 ng/mL EGF had a positive influence on the progression of maturation to MII at 72 h. The effect may not be related to the alteration of mRNA expression of genes analyzed in the present study, due to the complex patterns regulating meiotic arrest in canine oocytes. This work was supported by Grant no. 204119-03-1-LG000 from ARPC, Republic of Korea.

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