Project description:We previously showed that recombinant human chorionic gonadotropin (r-hCG) induces mammary gland differentiation and inhibits mammary tumorigenesis in rats. The present study investigates the impact of r-hCG on the stemness of mammary epithelial cells by using a rat model. We performed microarray analysis of RNA isolated from mammospheres formed by normal mammary epithelial cells extracted from rats treated for 21 days either with r-hCG or vehicle (control).
Project description:the aim of this work is to gain insight into Ovarianhyperstimulation Syndrome. All samples were mRNA from rat ovaries after 48 h of drug treatments.S1)1 IU PMSG + 1 IU hCG (this is the control)S2) 10 IU PMSG + 10 IU hCGS3) 40 IU PMSG + 30 IU hCG
Project description:Rat pituitary organ cultures were treated with either GnRH at 10nM for 6 hours, or treated with control for 6 h, both groups in triplicate; samples were processed to generate total RNA, which were subsequently analyzed for gene expression using Affymetrix rat 230 2.0 arrays Keywords: rat pituitary organ culture treated vs control
Project description:To further development of the effects of β-HCG in ovarian cancer SKOV3 cells,we have employed lncRNA and mRNA microarray as a discovery platform to identify lncRNA and mRNA expression in β-HCG overexpressing ovarian cancer cells. SKOV3 cells were transfected with lentiviral vector with eGFP, encoding β-HCG and negative control vector (LV- β-HCG and LV-CON,) by using polybrene. The dysregulation of β-HCG was confirmed by using RT-PCR. RNA was extracted and detected by a lncRNA and mRNA microarray in LV-β-HCG and LV-CON SKOV3 cells. The different expression of lncRNA and mRNA in LV-β-HCG and LV-CON SKOV3 cells was analyzed to explore the mechanism that β-HCG affect ovarain cancer cells.
Project description:The objective of this experiment was to explore differences in follicle transcriptomes in patients having oocyte maturation induced with either a bolus of hCG or a GnRH agonist. Upon oocyte retrieval, mural granuloasa cells and cumulus cells were isolated, allowing for comparison of the follicular transcriptomes between patients treated with eihter hCG or the GnHR agonist.
Project description:Gene expression profiles of sandwich-cutlured primary rat hepatocytes exposed to 5 mM and 10 mM acetaminophen were used in a parallelogram approach in order to compare gene expression responses between rat and human using in vitro cellular models, heaptocytes, and between rat in vitro and in vivo Experiment Overall Design: Samples were retrieved from acetaminophen treated rat hepatocyte cultures from 3 rats (3 biological replicates). Rat hepatocytes from each replicate were cultured in two culture conditions, each treated with 0, 5, and 10 mM acetaminophen for 24 h. This resulted in (3 rats x 2 culture conditions x 3 doses) 18 dual channel arrays on which control (0 mM acetaminophen) and reference samples (0, 5, and 10 mM acetaminophen) were hybridized
Project description:Cortical rat neuronal culture, lysis and proteolytic digestion Primary rat cortical neurons were generated from E18 Sprague-Dawley rat embryos with minor modifications (Swanger, Mattheyses et al. 2015, Henderson, Greathouse et al. 2019). Neurons were cultured at a density of 4x105 cells per well in 12-well culture plates (Fisher Scientific, catalog no. 353043). Neurons were cultured in Neurobasal medium (Fisher Scientific, catalog no. 21103-049) supplemented with B27 (Fisher Scientific, catalog no.17504-044). Culture maintenance included a half media change every 2-3 days. At DIV 14, neurons were either treated with 150 ng/mL recombinant NRN1 protein (Abcam, ab69755) or vehicle treated with diH2O for 6 hours. NRN1 concentration was chosen based on published data that identified a plateau in exogenous NRN1 induced effects on transient potassium currents at 150 ng/mL (Yao et al., 2012). After 6 hours neurons were washed 2x with 1 mL 1X phosphate-buffered saline (PBS). To harvest cells, 1 mL 1X PBS + protease inhibitor (Fisher Scientific, catalog no. 78426) was added and cells were centrifuged for 2300rpm for 5 minutes at 4°C. Cell pellets were lysed in 200uL 8M urea buffer and HALT protease and phosphatase inhibitor cocktail (1x final concentration). Lysates were sonicated with a probe sonicator 3 times for 10 s with 10 s intervals at 30% amplitude and cleared of cellular debris by centrifugation in a tabletop centrifuge at 18,000 rcf for 3 minutes at 4° C. Protein concentration was determined by BCA assay and one-dimensional SDS-PAGE gels were run followed by Coomassie blue staining as quality control for protein integrity and equal loading before proceeding to protein digestion. Protein homogenates (50 µg) were diluted with 50 mM NH4HCO3 to a final concentration of less than 2 M urea and then treated with 1 mM dithiothreitol (DTT) at 25°C for 30 minutes, followed by 5 mM iodoacetimide (IAA) at 25°C for 30 minutes in the dark. Protein was digested with 1:100 (w/w) lysyl endopeptidase (Wako) at 25°C for 2 hours and further digested overnight with 1:50 (w/w) trypsin (Pierce) at 25°C. Resulting peptides were desalted with a Sep-Pak C18 column (Waters) and dried under vacuum.
Project description:Influence of ovarian stimulation with 200 IU of hCG, (administered in the late follicular phase among ICSI patients undergoing a GnRH-antagonist protocol), on the endometrium on the day of oocyte pick-up. The purpose of the present study is to assess the influence of the administration of low dose hCG on the endometrium. In addition, by analysing the correlation of the morphological pattern and gene expression profile of human endometrium on the day of oocyte retrieval in patients of both treatment groups, we want to study the implantation potential. Keywords: gene expression analysis