scholarly journals Effects of Epidermal Growth Factor and Myo-Inositol on Nuclear Configuration and Subsequent Embryonic Development of Sheep Oocytes

2021 ◽  
Vol 9 (12) ◽  
pp. 2147-2152
Author(s):  
Omar Mardenli ◽  
Hadi Awad Hassooni ◽  
Mahdi Saleh Mohammad Alkerwi
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Culture Media ◽  
Calf Serum ◽  
Germinal Vesicle ◽  
Cellular Level ◽  
Biological Interactions ◽  
Epidermal Growth ◽  
Germinal Vesicle Break Down ◽  
Metaphase I

Growth factors and vitamin-like substances have great positive importance in most biological interactions in the cellular level. The addition of these elements in the culture media will increase the yield of the resulting embryos and improve quality. We examined the effects of epidermal growth factor (EGF) and myo-inositol (MI) on meiotic maturation and yields of blastocyst of Awassi sheep oocyte across two experiments. The oocytes obtained were subjected into three treatments: A (without EGF nor MI), B (10 ng/ml EGF + 20 mmol/l MI) and C (50 ng/ml EGF +40 mmol/l MI). Oocytes were then cultured in Ham's F-10 medium supplemented with 5% (v: v) fetal calf serum and 40 ng/ml follicle - stimulating hormone. In the first experiment, during the 27-h culture period, the oocytes were assessed for germinal vesicle break down, metaphase-I and metaphase-II stages across three-time intervals (9, 21 and 27-h). Results of the experiment showed that EGF and MI enhanced the rates of germinal vesicle break down phase (1.53%; 27-h interval; lowest value), metaphase-I (33.87%; 21-h interval) and metaphase-II (89.23%; 27-h interval). In the second experiment, the oocytes incubated in treatment B achieved the highest rates of cleavage (81.96%), 2-8 cell (62.35%) and blastocyst (45.09%). It is concluded from the present study that incubating sheep oocytes in culture media containing a cocktail of EGF (10 ng/ml) and MI (20 mmol/l) significantly improves the rates of metaphase-II, fertilization and blastocyst rates.

4-Hydroxyestradiol improves mouse embryo quality, epidermal growth factor-binding capability in vitro and implantation rates

2020 ◽  
Author(s):  
Nuria Hernández ◽  
Marta López-Morató ◽  
Mario J Perianes ◽  
Soledad Sánchez-Mateos ◽  
Vanessa Casas-Rua ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Embryo Quality ◽  
Culture Media ◽  
Embryo Implantation ◽  
In Vivo Models ◽  
Endometrial Cells ◽  
Epidermal Growth ◽  
Binding Capability

Abstract Embryo implantation in the uterus is a critical step to achieve success following ART. Despite favorable uterine conditions, a great number of good quality embryos fail to implant, often for reasons that are unknown. Hence, improving the implantation potential of embryos is a subject of great interest. 4-Hydroxyestradiol (4-OH-E2), a metabolic product of estradiol produced by endometrial cells, plays a key role in endometrial–embryonic interactions that are necessary for implantation. Nonetheless, the effects of 4-OH-E2 on embryos obtained in vitro have not been yet described. This study was designed to determine whether culture media enriched in 4-OH-E2 could improve the quality and implantation rate of embryos obtained in vitro, using both in vitro and in vivo models. We also analyzed its effects on the epidermal growth factor (EGF)-binding capability of the embryos. Our results showed that the presence of 4-OH-E2 in the culture media of embryos during the morula to blastocyst transition increases embryo quality and attachment to endometrial cells in vitro. 4-OH-E2 can also improve viable pregnancy rates of mouse embryos produced in vitro, reaching success rates that are similar to those from embryos obtained directly from the uterus. 4-OH-E2 improved the embryos’ ability to bind EGF, which could be responsible for the increased embryo implantation potential observed. Therefore, our results strongly suggest that 4-OH-E2 is a strong candidate molecule to supplement human IVF culture media in order to improve embryo implantation. However, further research is required before these findings can be translated with efficacy and safety to fertility clinics.

Proliferative response and oncogene expression induced by epidermal growth factor in EL2 rat fibroblasts

1986 ◽  
Vol 6 (6) ◽  
pp. 2275-2278
Author(s):  
E Liboi ◽  
E Pelosi ◽  
U Testa ◽  
C Peschle ◽  
G B Rossi
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Calf Serum ◽  
Fibroblast Growth ◽  
Fibroblast Line ◽  
Competence Factor ◽  
Step Model ◽  
Epidermal Growth ◽  
Rat Fibroblasts ◽  
Progression Factor

Extensive evidence supports a two-step model for the control of fibroblast growth, which includes first the action of a competence factor (e.g., platelet-derived growth factor) followed by the stimulus of a progression factor (e.g., epidermal growth factor [EGF]). We investigated whether this model may be applied to the euploid EL2 fibroblast line recently isolated from rat embryos (E. Liboi, M. Caruso, and C. Basilico, Mol. Cell. Biol. 4:2925-2928, 1984). Our results clearly show that EGF alone leads EL2 cells to proliferate in serum-free conditions at a rate corresponding to 50 to 60% of that observed in the presence of 10% calf serum. It is of interest that, when resting EL2 cells were exposed to EGF, transcription of both c-myc and c-fos was markedly induced. Altogether, these observations suggest that, in contrast with the model of fibroblast growth mentioned above, EL2 cells require the presence of a single growth factor (EGF) for induction of DNA synthesis, and the expression of myc and fos proto-oncogenes may represent an obligatory step in the pathway of commitment of EL2 cells to proliferation. In addition, we showed that EGF may induce EL2 cells to acquire some properties of transformed cells, such as growth in agar and loss of contact inhibition. This suggests that the particular response to EGF of the EL2 line may be strictly connected with the expression of a transformed phenotype.

143 EFFECT OF FSH OR EPIDERMAL-GROWTH-FACTOR-LIKE PEPTIDE SUPPLEMENTATION TO MATURATION MEDIUM ON DEVELOPMENTAL COMPETENCE OF BOVINE OOCYTES DERIVED FROM FULLY DEVELOPED FOLLICLES INDUCED BY SUPER-STIMULATION

2017 ◽  
Vol 29 (1) ◽  
pp. 180
Author(s):  
T. Yamanouchi ◽  
S. Sugimura ◽  
H. Matsuda ◽  
M. Ohtake ◽  
Y. Goto ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Formation Rate ◽  
Calf Serum ◽  
Developmental Competence ◽  
Blastocyst Formation ◽  
Maturation Medium ◽  
Epidermal Growth ◽  
Bovine Oocytes

Bovine oocytes obtained by ovum-pick-up (OPU) following follicle growth treatment (FGT) have improved quality and competence (Imai et al. 2008 Reprod. Fertil. Dev. 20, 182). However, the effect of the presence of FSH or epidermal growth factor (EGF) like peptide during in vitro maturation (IVM) on the developmental competence of FGT oocytes has not been well known. This study was undertaken to examine the developmental competence of FGT oocytes following IVM in the presence of FSH (recombinant human FSH) or EGF-like peptide (amphiregulin; Areg) and IVF. Japanese Black cows (n = 17) were used as donors. Five days after arbitrary OPU (opu group), follicles ≥8 mm in diameter were aspirated again, a controlled internal drug release (CIDR) was inserted into the vagina, and then pFSH was injected twice a day from the evening of Day 6 to the morning of Day 10 with decreasing doses (total of 20 AU; 4, 4, 3, 3, 2, 2, 1, 1 AU/day). On the evening of Day 8, PGF2α (0.5 mg of cloprostenol) was administered. On Day 11, oocytes were aspirated from follicles with ≥5 mm in diameter of the treated donors by OPU (fgt group). The cumulus-oocyte complexes (COC) were cultured in the absence (opu-cont and fgt-cont groups) or presence of 0.1 IU mL−1 FSH (opu-fsh and fgt-fsh groups) or 100 ng mL−1 Areg (opu-areg and fgt-areg groups) in IVM medium (mTCM199 containing 5 mg mL−1 BSA) for 20 to 22 h (1 COC/5 µL, total of 162–171 COC per group), and then co-cultured with 3 × 106 sperm/mL for 6 h. The presumptive zygotes were continued to culture in mCR1aa supplemented with 5% newborn calf serum for 216 h (1 zygote/5 µL) using micro-well culture dishes (Dai-Nippon-Print). When repeating this opu-fgt session in the same cow, an interval at least for 50 days was kept, and the session was performed 28 times. Statistical analysis was carried out by Mann-Whitney’s U-test (between opu and fgt groups) or Steel-Dwass test after Kruskal-Wallis test (among all groups). The number of follicles ≥5 mm increased in the fgt than opu group (17.8 v. 2.9; P < 0.01). The number of COC collected was not different between the opu and fgt groups (23.1 v. 19.6; P > 0.05). The blastocyst formation rate was higher in the fgt than opu group (36.9 v. 23.1%; P < 0.01). Within 6 groups, the blastocyst formation rate was higher in the fgt-fsh (43.3%; P < 0.01) and fgt-areg (39.5%; P < 0.05) groups than the opu-cont (16.3%) group. The rate in the fgt-fsh group was also higher than that in the opu-fsh group (43.3 v. 18.7%; P < 0.01). These results suggested that FGT improved the developmental competence of bovine oocytes, probably through improving the ability of the COC to react against FSH/Areg.

Somatomedin-like activity from cultured embryo-derived cells: partial characterization and stimulation of production by epidermal growth factor (urogastrone)

1984 ◽  
Vol 62 (12) ◽  
pp. 1335-1342 ◽  
Author(s):  
Paul R. Atkison ◽  
R. Marvin Bala ◽  
Morley D. Hollenberg
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cell Line ◽  
Culture Media ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
Placental Lactogen ◽  
Organ Cultures ◽  
Epidermal Growth ◽  
Stimulation Of

We have measured the appearance in cell culture media of a somatomedin-like activity (SLA) that cross-reacts in a basic-somatomedin radioreceptor assay and radioimmunoassay and that is produced by cultures of normal fetal-derived human (WI-38 fibroblasts) and mouse (embryonic palate) tissues and by cultures of a Simian virus 40 transformed WI-38 fibroblast cell line (WI-38 VA13/2RA subline). In addition to human growth hormone (hGH), epidermal growth factor (urogastrone) (EGF-URO), human placental lactogen (hPL), and porcine insulin (INS) were able to stimulate the production of SLA in serum-free cultures of the normal WI-38 fibroblasts; human prolactin (hPRL) caused only a marginal stimulation of SLA production in these cultures. In order of the magnitude of the stimulation of SLA production, the effects of the several hormones tested were [Formula: see text]. The half-maximal stimulatory effect of EGF-URO in the WI-38 cultures was observed at a concentration of about 1 nM. In the mouse palate organ cultures, EGF-URO also stimulated SLA production; the effect of EGF-URO was more pronounced in palate organ cultures derived from day 13 embryos compared with tissue obtained from embryos at days 14 and 15. The SLA produced by the human VA13/2RA subline was characterized further by gel filtration under acid conditions, by Procion dye column chromatography, and by high pressure liquid chromatography (HPLC). In terms of their immunoreactivity, adsorption to the dye column, and their apparent molecular weights (about 45 000), the SLA's produced by the VA13/2RA cells and the WI-38 cell cultures did not appear to differ. However, in terms of molecular weight and HPLC behaviour, the VA13/2RA-derived SLA was distinct from human basic somatomedin. We conclude that fetal-derived cells are capable of producing SLA in a manner that can be regulated both by a variety of polypeptide hormones and by the stage of embryonic development. Further, we conclude that the SLA produced by the fetal-derived human VA13/2RA cell line is indistinguishable from the SLA produced by WI-38 cells and that this SLA differs from the immunologically cross-reacting somatomedin that has been previously isolated from human plasma Cohn fraction.

scholarly journals pp60c-src tyrosine kinase, myristylation, and modulatory domains are required for enhanced mitogenic responsiveness to epidermal growth factor seen in cells overexpressing c-src.

1989 ◽  
Vol 9 (4) ◽  
pp. 1536-1544 ◽  
Author(s):  
L K Wilson ◽  
D K Luttrell ◽  
J T Parsons ◽  
S J Parsons
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Tyrosine Kinase ◽  
Cellular Proliferation ◽  
Control Cell ◽  
Calf Serum ◽  
Functional Requirements ◽  
Src Tyrosine Kinase ◽  
Clonal Cell Lines ◽  
Epidermal Growth

In previous studies examining the potential role of pp60c-src in cellular proliferation, we demonstrated that C3H10T1/2 murine embryo fibroblasts overexpressing transfected chicken genomic c-src displayed an epidermal growth factor (EGF)-induced mitogenic response which was 200 to 500% of the response exhibited by parental control cells (Luttrell et al., Mol. Cell. Biol. 8:497-501, 1988). In order to examine specific structural and functional requirements for pp60c-src in this event, 10T1/2 cells were transfected with chicken c-src genes encoding pp60c-src deficient in tyrosine kinase activity (pm430), myristylation, (pm2A), or a domain hypothesized to modulate the interaction with substrates or regulatory components (dl155). Neomycin-resistant clonal cell lines overexpressing each of the mutated c-src genes were assayed for EGF mitogenic responsiveness by measuring [3H]thymidine incorporation into acid-precipitable material or into labeled nuclei. The results were compared with those obtained with lines overexpressing the cDNA form of wild-type (wt) c-src or control cells transfected with the neomycin resistance gene only. As previously described for cells overexpressing wt genomic c-src (Luttrell et al., 1988), clones overexpressing wt cDNA c-src also exhibited enhanced EGF mitogenic responses ranging from approximately 300 to 400% of the control cell response. In contrast, clones overexpressing unmyristylated, modulation-defective, or kinase-deficient c-src not only failed to support an augmented response to EGF but also exhibited EGF responses lower than that of the control cells. Furthermore, there were no significant differences in the mitogenic responses to 10% fetal calf serum among any of the cells tested. These results indicate that pp60(c-scr) can potentiate mitogenic signaling generated by EGF but not all growth factors. This potentiation requires the utilization of pp60(c-scr) myristylation, and modulatory and tyrosine kinase domains and can me mediated by cDNA-encoded as well as by genome-encoded wt pp60(c-scr).

scholarly journals In-vitro maturation of human germinal vesicle stage oocytes: role of cumulus cells and epidermal growth factor in the culture medium

1998 ◽  
Vol 13 (6) ◽  
pp. 1638-1644 ◽  
Author(s):  
P. T. Goud ◽  
A. P. Goud ◽  
C. Qian ◽  
H. Laverge ◽  
J. Van der Elst ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Epidermal Growth

256 EFFECT OF EPIDERMAL GROWTH FACTOR ON IN VITRO MATURATION OF CYNOMOLGUS MONKEY (MACACA FASCICULARIS) OOCYTES

2008 ◽  
Vol 20 (1) ◽  
pp. 208
Author(s):  
J. Yamasaki ◽  
J. Okahara-Narita ◽  
C. Iwatani ◽  
H. Tsuchiya ◽  
S. Nakamura ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
In Vitro Maturation ◽  
Polar Body ◽  
Germinal Vesicle ◽  
Cynomolgus Monkeys ◽  
Feeder Layers ◽  
Maturation Rate ◽  
Epidermal Growth

Collected oocytes include not only mature oocytes (metaphase II: MII), but also immature oocytes (germinal vesicle: GV, and metaphase I: MI). To establish a dependable artificial indoor breeding program in cynomolgus monkeys, we are planning to carry out in vitro maturation (IVM) using GV and MI oocytes. In this study, we attempted to determine whether different types of feeder layers and epidermal growth factor (EGF) were effective for IVM. Cumulus–oocyte complexes (COCs) were collected from ovaries of 4–10-year-old female cynomolgus monkeys stimulated by the combination of FSH (25 IU kg–1 × 9 days) and hCG (400 IU kg–1) (Torii 2000 Primates 39, 399–406). Oocytes were classified by morphological features: oocytes retaining an intact germinal vesicle nucleus (GV); oocytes that had undergone germinal vesicle breakdown without polar body formation (MI); and oocytes with a first polar body (MII). GV and MI oocytes were co-cultured on monkey cumulus cells (MCC), monkey follicular ovarian cells (MFOC), monkey oviductal cells (MOC), or human solubilized amnion product (HSAP), with TCM-199+10% fetal bovine serum containing epidermal growth factor (EGF; 10 ng mL–1 or 20 ng mL–1). The maturation rate from GV to MII oocytes was 6.7% (MCC), 18.0% (MFOC), 35.7% (MOC), and 28.6% (HSAP) (Table 1). Although higher maturity was observed in MOC and HSAP, the effect of EGF was not found in co-cultures using any feeder layers. The maturation rate from MI to MII oocytes was 33.3% (MCC), 27.8% (MFOC), 55.6% (MOC), and 44.0% (HSAP) (Table 1). The highest maturation rate from GV and MI was observed in co-cultures using MOC. The maturation rate from MI to MII oocytes in the presence of 10 ng mL–1 EGF was 75.0% (MCC) and 73.7% (HSAP) (Table 1), whereas the rate in the presence of 20 ng mL–1 EGF was 59.1% (MCC), 64.3% (MFOC), 92.3% (MOC), and 60.0% (HSAP) (Table 1). Thus, the best maturation rate was a co-culture using MOC as a feeder layer with 20 ng mL–1 EGF. According to our results, maturation rate during IVM depends on the cellular type of feeder layers and the concentration of EGF. EGF is especially effective for maturity from MI to MII oocytes, but not from GV to MI or MII oocytes. Thus, IVM should be carred out under optimal culture conditions, including suitable feeder layer and media plus supplements. In the future, it is important that intracytoplasmic sperm injection be carried out using in vitro-matured MII oocytes for establishment of an artificial indoor breeding program in cynomolgus monkeys. Table 1. Number of matured oocytes co-cultured with different feeder layers and EGF

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