scholarly journals Mouse oocyte meiotic resumption and polar body extrusion in vitro are differentially influenced by FSH, epidermal growth factor and meiosis-activating sterol

2004 ◽  
Vol 19 (12) ◽  
pp. 2913-2918 ◽  
Author(s):  
G. Coticchio ◽  
G. Rossi ◽  
A. Borini ◽  
C. Grøndahl ◽  
G. Macchiarelli ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Polar Body ◽  
Mouse Oocyte ◽  
Epidermal Growth ◽  
Polar Body Extrusion

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

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.

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