ABSTRACT: The PANC-1 cells were maintained on DMEM supplemented with 10% FBS, 1% penicillin-streptomycin. ChIP-seq of PANC-1 cells using anti-H3K4me3Q5ser.
Project description:The PANC-1 cells were maintained on DMEM supplemented with 10% FBS, 1% penicillin-streptomycin. RNA-seq of PANC-1 cells after TGM2 KD.
Project description:In this study we analyzed the proteomic profiles of H460, and MCF7 3D tumor spheroids grown either in standard medium (3D_SM), or in FBSlow conditions (3D_FBSlow) and compared them to their relative 2D cultures (2D). Standard medium consisted of DMEM/F-12 supplemented with 0.5% Glucose, 2.5mM L-Glutamine, 2% B-27, 5μg/ml Heparin, 20μg/ml Insulin, 20ng/ml EGF, 20ng/ml Recombinant Human bFGF, 0.1% Bovine Serum Albumin (BSA) and 1% (v/v) of penicillin/streptomycin 100U/ml. FBSlow medium consisted of customized nutrient-restricted RPMI or DMEM culture media supplemented with only 2% FBS.
Project description:This study provides a comprehensive evaluation of changes in gene expression during treatment with environmental chemicals in embryonic bodies derived from mouse embryonic stem (ES) cells. ES cells were maintained under the feeder cells in phenol red-free DMEM Medium (supplemented with 15% FBS, 100uM Non-essential amino acids (NEAA), 1000U/ml Leukemia inhibitory factor(LIF), 100uM 2-mercaptoethanol(2-ME), and 0.5% penicillin/streptomycin). Cells were gently washed in warm PBS and transferred to phenol red-free DMEM Medium (supplemented with 15% KnockOut Serum Replacement, 100uM NEAA, 100uM 2-ME, and 0.5% penicillin/streptomycin) in Microsphere array (MSA300F) and exposed with low and high levels of each chemical just after removing LIF from the culture media. Prior to collection, cells were washed in warm PBS, resuspended and briefly incubated in the QIAGEN RLT buffer, and finally collected in triplicate at 48 hours. Following RNA isolation, the best RNA yields for each replicate set was selected for target preparation and microarray processing.
Project description:Analysis of differentially expressing genes in whole genome wide analysis of aptamer SQ2 positive cells (Capan-1, Panc-1, Panc-1+ve) and SQ2 negative cells (Panc-1-ve and HPDE) Panc-1 +ve and Panc-1-ve cell lines were generated from Panc-1 cell line based upon its hetergenous binding to aptamer SQ2. Detailed procedure of generation of these cell lines are described in Pooja Dua, Hye Suk Kang, Seung-Mo Hong, Ming-Sound Tsao, Soyoun Kim, and Dong-ki Lee. 2012 Alkaline Phosphatase ALPPL2 is a novel pancreatic carcinoma-associated protein. Cancer Research A five chip study using total RNA recovered from Capan-1, Panc-1, Panc-1+ve, Panc-1-ve and HPDE cells. Each chip measures 45,033 genes with three 60 mer probe pairs per target.
Project description:Investigation of DNA methlation changing in thymidine treatment-induced MEFs. Mouse embryonic fibroblasts (MEFs) were obtained at E14.5 embryos and supplied with Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen) with 15% FBS, 100 μg/ml streptomycin and 100 U/ml penicillin. Potential hematopoietic progenitor cells (CD45+CD41+CD31+c-Kit+) were negatively selected with Biotin labeled anti-CD45, anti-CD31, anti-CD41 and anti-c-Kit antibodies by magnetic beads. Hereafter, treated MEFs are referred to as 4neg MEFs for initiation of induction. 4neg MEFs were seeded on mitomycin (MMC)-treated MEFs as feeder cells and cultured in DMEM with 15% FBS, 100 μg/ml streptomycin, 100 U/ml penicillin, 2 mM L-glutamine, 0.1 mM non-essential amino acids and 100 μM thymidine as induction medium for 2 days followed with analysis
Project description:CMT-Stylo is a canine mammary gland adenocarcinoma cell line. From this cell line, a doxorubicin resistant subline called CMT-Star was developed by culturing the cells in the presence of doxorubicin from low concentration to 100 nM doxorubicin (to induce drug resistance). The CMT-Stylo cells were maintained in Dulbecco’s modified eagles medium (DMEM) supplemented with 10% Foetal bovine serum (FBS) and 1% Penicillin-Streptomycin (ThermoFisher Scientific, USA) in 5% CO2 at 370C. The CMT-Star cell line was maintained in additional 5 nM doxorubicin to maintain its resistant phenotype.
Project description:CMT-Stylo is a canine mammary gland adenocarcinoma cell line. From this cell line, a doxorubicin resistant subline called CMT-Star was developed by culturing the cells in the presence of doxorubicin from low concentration to 100 nM doxorubicin (to induce drug resistance). The CMT-Stylo cells were maintained in Dulbecco’s modified eagles medium (DMEM) supplemented with 10% Foetal bovine serum (FBS) and 1% Penicillin-Streptomycin (ThermoFisher Scientific, USA) in 5% CO2 at 370C. The CMT-Star cell line was maintained in additional 5 nM doxorubicin to maintain its resistant phenotype.
Project description:Purpose: To understand the molecular mechanisms underlying Cd exposure-induced diseases. Methods: Immortalized human bronchial epithelial cells (BEAS-2B) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Cellgro) supplemented with 1% Penicillin Streptomycin and 10% Fetal Bovine Serum (FBS, Atlanta Biologicals) at 37 degree C and 5 % CO2. For Cd exposure, 0, 2.5, 5 and 10 µM of CdCl2 was added to the media and the cells were cultured for 72h. Results: This study shows that cadmium exposure induces SNAIL1 expression via miR-30e downregulation and the cells undergo epithelial-mesenchymal transition.
Project description:Investigation of differentially expressed genes in thymidine treatment-induced MEFs. Mouse embryonic fibroblasts (MEFs) were obtained at E14.5 embryos and supplied with Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen) with 15% FBS, 100 μg/ml streptomycin and 100 U/ml penicillin. Potential hematopoietic progenitor cells (CD45+CD41+CD31+c-Kit+) were negatively selected with Biotin labeled anti-CD45, anti-CD31, anti-CD41 and anti-c-Kit antibodies by magnetic beads. Hereafter, treated MEFs are referred to as 4neg MEFs for initiation of induction. 4neg MEFs were seeded on mitomycin (MMC)-treated MEFs as feeder cells and cultured in DMEM with 15% FBS, 100 μg/ml streptomycin, 100 U/ml penicillin, 2 mM L-glutamine, 0.1 mM non-essential amino acids and 100 μM thymidine as induction medium for 6 days. Cells were then trypsinized and re-seeded to new feeder cells for another 6 days. Cells were positively selected with Biotin labeled anti-CD45 by magnetic beads followed with analysis.
Project description:The goal of the study was to identify genes and pathways that were altered when human pancreatic ductal adenocarcinoma (PDAC) cancer cells are cultured with different carbon source (Glucose versus Galactose). Primary adherent cultures established from patient-derived xenograft passaged in mice were established (PancA6L). Low passage (< 15) PDX-derived primary PDAC PancA6L cultures were trypsinized and seeded at a concentration of 800,000 cells in p100 plates with RPMI medium supplemented with 10% fetal bovine serum (FBS) and 50 units/mL of penicillin and streptomycin. After 24 h, cells were cultured with either 1) glucose-free DMEM medium (Dulbecco´s Modified Eagle Medium, Thermo Fisher Scientific) supplemented with 5mM glucose (0.9 g/L), 10% FBS, 50 units/mL of penicillin and streptomycin and 1mM of pyruvate [Glucose: OXPHOS-independent conditions] or 2) glucose-free DMEM medium (Thermo Fisher Scientific) supplemented with 5mM galactose (0.9 g/L), 10% FBS, 50 units/mL of penicillin and streptomycin and 1mM of pyruvate [Galactose: OXPHOS-competent enriched conditions]. Sugar concentrations of 5mM were chosen to mimic physiological sugar levels (glucose, 5mM) and to avoid potential biological artifacts mediated by supraphysiological sugar levels. Media for both conditions were changed every day Following 14 days in culture as spheres, Total RNA was isolated by the guanidine thiocyanate (GTC) method using standard protocols. PolyA+ RNA fraction was processed as in Illumina’s ‘‘TruSeq RNA Sample Preparation v2 Protocol’’. The resulting purified cDNA library was applied to an Illumina flow cell for cluster generation (TruSeq cluster generation kit v5) and sequenced on the Genome Analyzer IIx with SBS TruSeq v5 reagents by following manufacturer’s protocols. RNA-seq data sets were analyzed using the tool Nextpresso.