328 EPIDERMAL GROWTH FACTOR ENHANCES MEIOTIC MATURATION OF CANINE OOCYTES IN THE PRESENCE OF BSA

2006 ◽  
Vol 18 (2) ◽  
pp. 271
Author(s):  
Y.-X. Jin ◽  
X.-H. Shen ◽  
X.-S. Cui ◽  
I.-K. Kong ◽  
N.-H. Kim
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Estrous Cycle ◽  
Oocyte Quality ◽  
Culture Conditions ◽  
Meiotic Maturation ◽  
Lower Percentage ◽  
North Carolina State University ◽  
Epidermal Growth

Despite many attempts to improve the in vitro maturation (IVM) of canine oocytes using various culture conditions, the efficiency of canine IVM remains very low compared with that of other domestic animals. In the present study we examined the effect of (1) ovarian estrous stage on oocyte quality, and (2) epidermal growth factor (EGF) in the presence and absence of macromolecules on the IVM of canine oocytes. Reproductive tracts from normal, client-owned bitches aged approximately 7 months were collected after routine ovariohysterectomy at private veterinary clinics. The ovaries were removed, washed free of blood with PBS, and repeatedly cut with a scalpel blade at room temperature in HEPES-buffered Tyrodes medium containing 0.1% (w/v) polyvinyl alcohol. Each group of 10 cumulus-oocyte complexes (COCs) was matured in North Carolina State University (NCSU)-17 medium supplemented with 1 �g/mL FSH, 10 IU/mL hCG, and 1 �g/mL �-estradiol under paraffin oil for 48 or 72 h at 37�C. Follicular ovaries contained the largest number of oocytes (more than 300), and the number of oocytes e100 �m in diameter was also significantly greater in follicular ovaries than in ovaries at other stages (P < 0.001). Luteal ovaries had the smallest number of both total and large oocytes. After 72 h of culture, significantly more oocytes recovered from follicular ovaries were in GVBD than oocytes from anestrous or luteal ovaries. After 72 h of culture, oocytes recovered from follicular ovaries had a significantly lower percentage reaching the GV stage (59.7 � 5.2%) and more in GVBD (40.3 � 3.1%) compared with both anestrous (GV, 75.1 � 5.2%; GVBD, 24.9 � 3.1%) and luteal (GV, 79.8 � 5.2%; GVBD, 22.0 � 3.1%) ovaries (P < 0.05). However, maturation to MI/MII was not statistically different between the stages of the estrous cycle. BSA or FBS supplementation did not improve meiotic maturation when compared to PVA supplementation. In the presence of 100 nM epidermal growth factor (EGF), either FBS or BSA enhanced the proportion of oocytes in GVBD/MI (59.7 � 2.2% [FBS] and 51.7 � 2.2% [BSA] vs. 36.1 � 2.2% [PVA], P < 0.05). In addition, 0.4% BSA significantly increased the percentage of oocytes in MII (18.5 � 0.8%) compared to 10% FBS (7.1 � 0.8%; P < 0.05). These results suggest that the estrous cycle of bitches influences the meiotic maturation of oocytes cultured in vitro, and EGF increases the meiotic maturation of canine oocytes in the presence of BSA in vitro. This work was funded by a grant from the Ministry of Science and Technology of Korea and the National Research Laboratory Program.

scholarly journals Epidermal Growth Factor Family Members: Endogenous Mediators of the Ovulatory Response

Endocrinology ◽  
2005 ◽  
Vol 146 (1) ◽  
pp. 77-84 ◽  
Author(s):  
H. Ashkenazi ◽  
X. Cao ◽  
S. Motola ◽  
M. Popliker ◽  
M. Conti ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Oocyte Maturation ◽  
Egf Receptor ◽  
Kinase Inhibitor ◽  
Meiotic Maturation ◽  
Expression Of Genes ◽  
Epidermal Growth ◽  
Stimulation Of

Previous studies showed that epidermal growth factor (EGF) and TGFα mimic the action of LH on the resumption of oocyte maturation. We tested whether EGF-like agents, such as amphiregulin (AR), epiregulin (ER), and betacellulin (BTC), also mediate the LH stimulation of the ovulatory response in the rat. LH induced transient follicular expression of AR, ER, and BTC mRNA, reaching a maximum after 3-h incubation. Furthermore, the addition of ER, AR, and BTC to the culture medium could mimic some of LH actions. AR and ER fully simulated LH-induced resumption of meiosis in vitro, whereas BTC was less effective. To study the putative involvement of EGF-like factors in mediation of LH signal, the effect of the EGF receptor kinase inhibitor AG1478 was tested. When added with LH, AG1478, but not its inactive analog AG43, reduced EGF receptor phosphorylation and oocyte maturation compared with follicles treated with LH only. In addition to the inhibition of resumption of meiosis, AG1478 administration into the bursa (3 μg/bursa) resulted in 51% (P &lt; 0.0005) inhibition of ovulation in the treated ovaries, compared with the untreated contralateral ones, as well as to the vehicle-treated ovaries (P &lt; 0.02). LH, as well as ER, induced the expression of genes associated with the ovulatory response like rat hyaluronan synthase-2, cycloxygenase-2, and TNFα-stimulated gene 6 mRNA, whereas AG1478 inhibited this effect of LH. Release of EGF-like factors from the membrane is dependent on activated metalloproteases. Indeed, Galardin, a broad-spectrum metalloprotease inhibitor, but not a specific matrix metalloprotease 2 and 9 inhibitor, suppressed meiotic maturation induced by LH. Conversely, meiotic maturation induced by ER was not affected by Galardin, thus, supporting the notion that LH releases follicular membrane-bound EGF-like agents. In summary, EGF-like factors such as ER, AR, and BTC seem to mediate, at least partially, the LH stimulation of oocyte maturation, ovulatory enzyme expression, and ovulation.

Effect of epidermal growth factor on the cumulus expansion, meiotic maturation and development of buffalo oocytes in vitro

1999 ◽  
Vol 144 (10) ◽  
pp. 266-267 ◽  
Author(s):  
S. Chauhan ◽  
S. K. Singla ◽  
P. Palta ◽  
R. S. Manik ◽  
M. L. Madan
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Meiotic Maturation ◽  
Cumulus Expansion ◽  
Epidermal Growth

scholarly journals Improvement of developmental competence of aged porcine oocytes by means of the synergistic effect of insulin-like growth factor-1 (IGF-1) and epidermal growth factor (EGF)

2011 ◽  
Vol 50 (No. 7) ◽  
pp. 300-310 ◽  
Author(s):  
I. Petrová ◽  
R. Rajmon ◽  
M. Sedmíková ◽  
Z. Kuthanová ◽  
F. Jílek ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Culture Conditions ◽  
Developmental Competence ◽  
Insulin Like Growth Factor ◽  
Large Numbers ◽  
Complex Process ◽  
Epidermal Growth ◽  
The Many

There is a constant increase in demand for large numbers of high-quality oocytes of domestic mammals for various reproductive biotechnologies. Prolonged in vitro culture represents one of the many possible ways of meeting this demand. However, oocytes further cultured in vitro after reaching the stage of metaphase&nbsp;II undergo the complex process called &ldquo;ageing&rdquo;, which decreases the quality of oocytes. The aim of the present study was to improve the culture conditions for in vitro matured porcine oocytes by supplement of the epidermal growth factor (EGF) or the insulin-like growth factor 1 (IGF-1) in order to reduce the adverse effects of ageing. Supplement of either EGF or IGF-1 had no significant effect on the ageing of porcine oocytes. Significant effects were demonstrated when the oocytes were cultured with both EGF and IGF-1. A combination of 10 &micro;g EGF/ml with 25 ng IGF-1/ml was the most effective. Oocytes aged under these conditions retained very good developmental competence. We observed development to the morula (21%) or blastocyst (25%) stage in oocytes aged for 1 day with EGF and IGF-1. Porcine oocytes matured in vitro are more resistant to the ageing when cultured in the presence of both EGF and IGF-1 and these conditions retain an elevated developmental competence for a certain time. &nbsp;

Computational Evaluation and In Vitro Validation of New Epidermal Growth Factor Receptor Inhibitors

2020 ◽  
Vol 20 (18) ◽  
pp. 1628-1639
Author(s):  
Sergi Gómez-Ganau ◽  
Josefa Castillo ◽  
Andrés Cervantes ◽  
Jesus Vicente de Julián-Ortiz ◽  
Rafael Gozalbes
Keyword(s):  
Cell Proliferation ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Binding Affinity ◽  
Cell Lines ◽  
Growth Factor Receptor ◽  
Significant Activity ◽  
Epidermal Growth

Background: The Epidermal Growth Factor Receptor (EGFR) is a transmembrane protein that acts as a receptor of extracellular protein ligands of the epidermal growth factor (EGF/ErbB) family. It has been shown that EGFR is overexpressed by many tumours and correlates with poor prognosis. Therefore, EGFR can be considered as a very interesting therapeutic target for the treatment of a large variety of cancers such as lung, ovarian, endometrial, gastric, bladder and breast cancers, cervical adenocarcinoma, malignant melanoma and glioblastoma. Methods: We have followed a structure-based virtual screening (SBVS) procedure with a library composed of several commercial collections of chemicals (615,462 compounds in total) and the 3D structure of EGFR obtained from the Protein Data Bank (PDB code: 1M17). The docking results from this campaign were then ranked according to the theoretical binding affinity of these molecules to EGFR, and compared with the binding affinity of erlotinib, a well-known EGFR inhibitor. A total of 23 top-rated commercial compounds displaying potential binding affinities similar or even better than erlotinib were selected for experimental evaluation. In vitro assays in different cell lines were performed. A preliminary test was carried out with a simple and standard quick cell proliferation assay kit, and six compounds showed significant activity when compared to positive control. Then, viability and cell proliferation of these compounds were further tested using a protocol based on propidium iodide (PI) and flow cytometry in HCT116, Caco-2 and H358 cell lines. Results: The whole six compounds displayed good effects when compared with erlotinib at 30 μM. When reducing the concentration to 10μM, the activity of the 6 compounds depends on the cell line used: the six compounds showed inhibitory activity with HCT116, two compounds showed inhibition with Caco-2, and three compounds showed inhibitory effects with H358. At 2 μM, one compound showed inhibiting effects close to those from erlotinib. Conclusion: Therefore, these compounds could be considered as potential primary hits, acting as promising starting points to expand the therapeutic options against a wide range of cancers.

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.

Sign in / Sign up

Export Citation Format

Share Document