ChIP H3K27me3 Mouse Canine

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - Cancer cell lines Leukemia cancer cell lines KG-1

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - Cancer cell lines Leukemia cancer cell lines THP-1

Get tips on using Human Breast Cancer Susceptibility Protein 2 (BRCA2) ELISA Kit to perform ELISA Human - BRCA2

Get tips on using Ion AmpliSeq™ Cancer Hotspot Panel v2 to perform Cell line authentication Lung carcinoma A549 cell line

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - Cancer cell lines Liver cancer cell lines Hepato cellular carcenoma (SMMC-7721, Huh7 & HepG2))

Get tips on using p21 Waf1/Cip1 (12D1) Rabbit mAb #2947 to perform Western blotting p21

Get tips on using Phusion Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Human - Deletion MCF-7 sodium channel β1 subunit

Get tips on using Phusion Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Human - Deletion MDA-MB-231 sodium channel β1 subunit

Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) have been greatly used for studies on embryonic development and cell differentiation.iPSCs provide a stable source for either self-renewal or differentiation into suitable cells when cultured in a particular environment. Pluripotent cell culture was originally started by deriving cells from inner cell mass (ICM) from pre-implanted blastocysts, these were called embryonic stem cells. These cells after isolation can be grown on traditional extracellular matrices (like mouse embryonic fibroblasts, MEFs) or feeder-free culture systems. DMEM/F12 has been the most commonly used basal media in the culture of pluripotent cells. These cells are cultured at normal atmospheric oxygen levels, 21%, however, some studies have proposed that 4% oxygen tension may be better for hESC growth. Higher D-glucose concentration (4.2g/l) and osmolarity (320mOsm) that mimics the natural environment of embryonic tissue are optimal for the growth of hESCs. Supplements like N2 and/or B-27, in the presence of growth factors like bFGF, have been shown to increase pluripotency of these cells. bFGF, FGF2 and other ligands of receptor tyrosine kinases like IGF are also required or maintain self-renewal ability of these cells. TGF𝛃1, by its activation of SMAD2/3 signalling, also represses differentiation of iPSCs. Other compounds like ROCK inhibitors reduce blebbing and apoptosis in these cells to maintain their clonogenicity. However, an inhibitor for LIF (leukaemia inhibitory factor, which is one of the pluripotent genes) has an opposing effect. Therefore, it is important to understand the culture conditions and media composition that affect downstream signalling in hESCs or iPSCs that may lead to their differentiation.