scholarly journals Hepatoma-derived growth factor: from the bovine uterus to the in vitro embryo culture

Reproduction ◽  
2014 ◽  
Vol 148 (4) ◽  
pp. 353-365 ◽  
Author(s):  
E Gómez ◽  
E Correia-Álvarez ◽  
J N Caamaño ◽  
C Díez ◽  
S Carrocera ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Cell Nuclei ◽  
Blastocyst Development ◽  
Fetal Fibroblast ◽  
Cell Counts ◽  
Apical Cytoplasm ◽  
Uterine Fluid ◽  
Protein Free Medium

Early in cow embryo development, hepatoma-derived growth factor (HDGF) is detectable in uterine fluid. The origin of HDGF in maternal tissues is unknown, as is the effect of the induction on developing embryos. Herein, we analyze HDGF expression in day 8 endometrium exposed to embryos, as well as the effects of recombinant HDGF (rHDGF) on embryo growth. Exposure to embryos did not alter endometrial levels of HDGF mRNA or protein. HDGF protein localized to cell nuclei in the luminal epithelium and superficial glands and to the apical cytoplasm in deep glands. After uterine passage, levels of embryonic HDGF mRNA decreased and HDGF protein was detected only in the trophectoderm. In fetal fibroblast cultures, addition of rHDGF promoted cell proliferation. In experiments with group cultures of morulae in protein-free medium containing polyvinyl alcohol, adding rHDGF inhibited blastocyst development and did not affect cell counts when the morulae were early (day 5), whereas it enhanced blastocyst development and increased cell counts when the morulae were compact (day 6). In cultures of individual day 6 morulae, adding rHDGF promoted blastocyst development and increased cell counts. Our experiments with rHDGF indicate that the growth factor stimulates embryonic development and cell proliferation. HDGF is synthesized similarly by the endometrium and embryo, and it may exert embryotropic effects by autocrine and/or paracrine mechanisms.

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.

scholarly journals Cell-free fat extract promotes tissue regeneration in a tissue expansion model

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Mingwu Deng ◽  
Xiangsheng Wang ◽  
Ziyou Yu ◽  
Yizuo Cai ◽  
Wei Liu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Collagen Type ◽  
Growth Factor Receptor ◽  
Tissue Expansion ◽  
Nuclear Antigen ◽  
Hacat Cells ◽  
Expansion Model

Abstract Background Tissue expansion techniques play an important role in plastic surgery. How to improve the quality of the expanded skin and shorten the expansion period are still worth investigating. Our previous studies found that a cell-free fat extract (CEFFE) possessed pro-angiogenic and pro-proliferative activities. However, the role of CEFFE on tissue expansion has remained unclear. The purpose of this study was to evaluate the effect of CEFFE on tissue expansion. Methods A rat tissue expansion model was used. Animals were treated with CEFFE by subcutaneous injection. After 4 weeks of tissue expansion, the skin necrosis and retraction rates were evaluated, the thicknesses of the epidermis and dermis were determined by histological analyses, blood vessel density was measured by anti-CD31 staining, cell proliferation was assessed by proliferating cell nuclear antigen staining, and the expression of specific proteins was evaluated by western blot analyses. In addition, the effects of CEFFE on the proliferation and cell cycle of cultured HaCaT cells were evaluated in vitro. Results CEFFE treatment significantly decreased the necrosis rate and retraction of the expanded skin. The thickness of the epidermal and dermal layers was higher in CEFFE-treated compared to untreated skin. The density of blood vessels and cell proliferation in the epidermis of the expanded skin was improved by CEFFE treatment. In addition, CEFFE treatment significantly increased the expression of the vascular endothelial growth factor receptor, epidermal growth factor receptor, collagen type 1, and collagen type 3. CEFFE also increased the proliferation of HaCaT cells in culture. Conclusions CEFFE improves the quality of the expanded skin by promoting angiogenesis and cell proliferation. It could be potentially used clinically for augmenting tissue expansion.

Differential effects of epidermal growth factor (EGF) on cell locomotion and cell proliferation in a cloned human embryonal carcinoma-derived cell line in vitro

1986 ◽  
Vol 86 (1) ◽  
pp. 47-55
Author(s):  
W. Engstrom
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cell Line ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Colony Diameter ◽  
Cell Locomotion ◽  
Epidermal Growth

The effects of epidermal growth factor (EGF) on clones from a human embryonal carcinoma-derived cell line (Tera-2) have been studied. Cells were plated at clonal densities, whereafter the effects of serum and EGF on cell locomotion and cell proliferation were examined. The addition of 50 ngEGF ml-1 resulted in increased migration, as judged by increased colony diameter in the presence of EGF. However, the effect of EGF on cell locomotion was rarely accompanied by any effect on cell proliferation. It was concluded that EGF exerts a preferential effect on cell migration in human embryonal carcinoma cells in vitro.

scholarly journals P460 Evaluation of potential mechanisms underlying the safety observations of filgotinib in clinical studies in rheumatoid arthritis

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S409-S409
Author(s):  
A Clarke ◽  
J Di Paolo ◽  
B Downie ◽  
A Meng ◽  
N Mollova ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Cell Proliferation ◽  
Clinical Studies ◽  
Nk Cell ◽  
Janus Kinase ◽  
Jak Inhibitor ◽  
Jak Inhibitors ◽  
Erythroid Progenitor ◽  
Cell Counts

Abstract Background Inhibitors of the Janus kinase-signal transducers and activators of transcription (JAK-STAT) pathway have demonstrated efficacy in the treatment of rheumatoid arthritis (RA) and inflammatory bowel disease (IBD). Differences in selectivity of JAK inhibitors for JAK1, JAK2, JAK3 and TYK2 may influence their respective safety profiles, and the mechanisms responsible are not currently known. Filgotinib (FIL), a JAK1 inhibitor, did not negatively impact haemoglobin, LDL:HDL ratios or natural killer (NK) cell counts in clinical trials. Here, we compare the in vitro mechanistic profiles of four JAK inhibitors at clinically relevant doses. Methods JAK inhibitors (FIL, FIL metabolite [GS-829845], baricitinib [BARI], tofacitinib [TOFA], and upadacitinib [UPA]) were evaluated in vitro in human-cell-based assays. Growth of erythroid progenitors from human cord blood CD34+ cells was assessed using a HemaTox™ liquid expansion assay, NK cell proliferation was induced by IL-15 and LXR agonist-induced cholesteryl ester transfer protein (CETP) expression was assessed in the hepatic cell line, HepG2. Using assay-generated IC50 values and the reported human plasma concentrations from clinical studies, we calculated the target coverage for each JAK inhibitor at clinically relevant doses. The activity of FIL in humans was based on PK/PD modelling of FIL + GS-829845. Results Inhibition of cellular activity was calculated for each JAK inhibitor based on in vitro dose-response data, human exposure data and modelled PK/PD relationships. At clinically relevant doses, FIL resulted in lower calculated inhibition of NK cell proliferation compared with other JAK inhibitors. FIL 100 mg and 200 mg also reduced CETP expression, whereas other JAK inhibitors had no effect. There was no difference in the effect of FIL vs. other JAK inhibitors on erythroid progenitor cell differentiation or maturation. Conclusion FIL, a JAK1 inhibitor, resulted in less inhibition of NK cell proliferation compared with BARI, TOFA, and UPA. FIL also reduced LXR agonist-induced CETP expression, while the other inhibitors did not alter these levels. These results provide a potential mechanistic link between the observed reduction of CETP concentration following FIL treatment and the previously observed reduction in the LDL:HDL ratio in RA patients.

scholarly journals miR-202 Inhibits Cell Proliferation, Migration, and Invasion by Targeting Epidermal Growth Factor Receptor in Human Bladder Cancer

2018 ◽  
Vol 26 (6) ◽  
pp. 949-957 ◽  
Author(s):  
Liqing Zhang ◽  
Jianjiang Xu ◽  
Gaodi Yang ◽  
Heng Li ◽  
Xiuxia Guo
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Growth Factor Receptor ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Epidermal Growth

Recent studies have demonstrated that miR-202 is associated with several types of cancer; however, the expression and function of miR-202 have not been investigated in bladder cancer. We analyzed the expression of miR-202 in bladder cancer tissues and adjacent noncancerous tissues. The effect of miR-202 on the proliferation, migration, and invasion was evaluated by in vitro assays. The target gene of miR-202 was assessed by luciferase reporter assay. In this study, miR-202 was found to be significantly downregulated in bladder cancer cell lines and tissues and was highly correlated with the T classification, N classification, grade, and recurrence. Ectopic expression of miR-202 suppressed cell viability, colony formation, cell migration, and invasion in vitro and inhibited xenograft tumor growth in vivo. Inversely, downregulation of miR-202 had contradictory effects. The 3′-untranslated region (3′-UTR) of epidermal growth factor receptor (EGFR) was identified as a direct target of miR-202 using luciferase reporter assays, and knockdown of EGFR enhanced miR-202-inhibited cell proliferation, migration, and invasion. In conclusion, miR-202 suppresses bladder cancer carcinogenesis and progression by targeting EGFR, thereby representing a potential target for miRNA-based therapy for bladder cancer in the future.

207 IN VITRO MATURATION TREATMENT AFFECTS DEVELOPMENTAL COMPETENCE OF LAPAROSCOPIC OVUM PICKUP-DERIVED OOCYTES IN FOLLICLESTIMULATING HORMONE-STIMULATED GOATS

2008 ◽  
Vol 20 (1) ◽  
pp. 182 ◽  
Author(s):  
Y. Locatelli ◽  
N. Poulin ◽  
G. Baril ◽  
J.-L. Touzé ◽  
A. Fatet ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Follicular Fluid ◽  
Developmental Competence ◽  
Postmortem Changes ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Development Rates ◽  
Epidermal Growth

The aim of the present study was to assess the effect of IVM treatment on the developmental competence of oocytes recovered from repeated laparoscopic ovum pickukp (LOPU) in goats. A total of 94 LOPU sessions were performed on 33 adult goats of the Saanen and Alpine breeds. Females were synchronized (Day 0) during the nonbreeding season by inserting vaginal sponges (45 mg of fluorogestone acetate, Intervet, Boxmeer, The Netherlands). At Day 8, an i.m. injection of 50 μg of cloprostenol (Estrumate; Schering-Plough Animal Health, Pointe-Claire, Quebec, Canada) was administered. Porcine FSH (Stimufol, Merial, Brussels, Belgium, 160 mg/goat) was administered in 5 injections at 12-h intervals, starting on Day 8. The LOPU took place under general anesthesia on Day 11, and follicles ≥2 mm were aspirated with an 18-gauge needle connected to a controlled vacuum system. Vaginal sponges were removed at the time of LOPU. Treatments were repeated 2 times in a 2-week interval scheme (2 goats and 1 goat were excluded from the experiment during the second and third LOPU sessions, respectively). Cumulus–oocyte complexes were washed and evaluated for quality (graded from 1 to 3). Oocytes recovered from unstimulated slaughterhouse-derived ovaries served as a control. Cumulus–oocytes complexes from Grades 1 and 2 were submitted to IVM in TCM-199, supplemented with 100 μm of cysteamine and either 10 ng mL–1 of epidermal growth factor (EGF) or 10% follicular fluid and 100 ng mL–1 of ovine FSH (FF-FSH). Matured oocytes were then submitted to IVF and in vitro development as described by Cognié et al. (2004 Reprod. Fertil. Dev. 16, 437–445). Over the 94 LOPU sessions, 20.4 ± 0.9 follicles were aspirated (mean ± SEM), allowing the recovery of 12.3 ± 0.7 COC per goat and per session, of which 80.1% were suitable for IVM (Grades 1 and 2). Results of in vitro production are detailed in the table. The IVM treatment did not significantly affect cleavage or blastocyst development rates in oocytes derived from slaughterhouse ovaries. Cleavage rates were significantly decreased in LOPU-derived oocytes when compared with control oocytes. For LOPU-derived oocytes, cleavage and final blastocyst development rates were increased significantly and kinetics of embryo development were accelerated when FF-FSH was used during IVM as compared with EGF. The IVM with FF-FSH allowed us to produce 4.1 blatocysts per goat per LOPU session. These results demonstrate the interest in LOPU for goat embryo production once appropriate IVM treatment is used. The difference observed between LOPU and slaughterhouse oocytes in terms of response to IVM treatments may be related to FSH stimulation prior to the LOPU session or to postmortem changes in oocyte responsiveness in the slaughterhouse group. Table 1. Effects of oocyte origin [laparoscopic ovum pickukp (LOPU) or slaughterhouse derived] and maturation treatment [epidermal growth factor (EGF) or follicular fluid (FF)-FSH] on in vitro embryo production (6 replicates)

92 INSULIN-LIKE GROWTH FACTOR-1 PROTECTS BOVINE PRE-IMPLANTATION EMBRYOS PRODUCED IN VITRO FROM ANTI-DEVELOPMENTAL ACTIONS OF MENADIONE

2016 ◽  
Vol 28 (2) ◽  
pp. 176
Author(s):  
N. A. S. Rocha-Frigoni ◽  
B. C. S. Leão ◽  
P. C. Dall'Acqua ◽  
M. Ambrogi ◽  
G. Z. Mingoti
Keyword(s):  
Oxidative Stress ◽  
Growth Factor ◽  
Molecular Probes ◽  
Insulin Like Growth Factor ◽  
Blastocyst Development ◽  
Chi Square ◽  
Fluorescent Intensity ◽  
Chi Square Test ◽  
Student’S T

The objective of this study was to evaluate the protective effect of insulin-like growth factor (IGF-1) on blastocyst development and cryotolerance of bovine embryos in in vitro culture (IVC) under oxidative stress induced by menadione (MD). Cumulus-oocyte complexes (n = 1421) were matured in TCM-199 with bicarbonate, hormones, and 10% FCS for 22 h. After fertilization, the presumptive zygotes were cultured up to 7 days in SOF medium with 2.5% FCS and 0.5% BSA (control), and also supplemented with 100 μM IGF-1 (IGF). At Day 6, MD was included in the culture medium (0 μM, control; or 5.0 μM, MD) during 24 h. Cultures were conducted at 38.5°C in 5% CO2 in air. The cleavage and blastocysts rates were evaluated, respectively, at Days 3 and 7 (IVF = Day 0). At Day 7, a sample of the blastocysts was stained with 5 μM H2DCFDA (Molecular Probes, Canada) to evaluate the intracellular ROS levels or was stained for TUNEL (In Situ Cell Death Detection Kit, Roche, Indianapolis, IN, USA). Stained embryos were immediately evaluated under an epifluorescence microscope (excitation 495/550 nm and emission 404/590 nm, respectively, for ROS and TUNEL), and the images of embryos stained with H2DCFDA were analysed by Q-Capture Pro image software for determining the fluorescent intensity. Other blastocysts were vitrified (Ingámed®, Maringá-PR, Brazil), and after warming, they were cultured for 24 h to evaluate the re-expansion rates. The results were compared by ANOVA followed by Student’s t-test (mean ± s.e.M) and re-expansion rates by chi-square test (P < 0.05). The cleavage rates did not differ (P > 0.05) among groups (77.1 ± 1.9% to 82.75 ± 2.2%). The blastocyst rates were similar between control (35.4 ± 2.0%) and IGF (34.5 ± 3.7%), and both were higher (P < 0.05) than MD (21.3 ± 2.7%); the IGF+MD group (28.3 ± 1.6%) was similar (P > 0.05) to all groups. The intracellular levels of ROS were higher (P < 0.05) for the MD group (21.7 ± 0.7) than for control (17.0 ± 1.6), and both were similar (P > 0.05) to the IGF (19.2 ± 0.6) and IGF+MD (18.0 ± 1.0) groups. The highest rates of apoptosis were found in the MD group (22.3% ± 2.3) and the smallest in IGF (9.1% ± 0.7), and both differed (P < 0.05) from control (12.8% ± 1.0), and IGF+MD (15.6% ± 1.6). The re-expansion rates were similar between control (77.4%) and IGF (69.2%), and both were higher (P < 0.05) than MD (49.1%); however, the IGF+MD group (57.6%) was similar (P > 0.05) to IGF and MD groups. In conclusion, the supplementation with IGF-1 during IVC reversed the detrimental effects of MD on embryonic levels of ROS and apoptosis, as well as improved the embryo development and cryotolerance of blastocysts under oxidative stress. Financial support was provided by FAPESP (#2012/10083–8 and #2013/07382–6).

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