ChIP acH3 Bovine Donkey

Get tips on using Gibco™DMEM, low glucose, pyruvate to perform Stem cell culture media Human bone mesenchymal stem cell (BMSC)

Get tips on using LC3B (D11) XP® Rabbit mAb to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)

Get tips on using CYTO-ID® Autophagy detection kit to perform Autophagy assay cell type - U2OS (human bone osteosarcoma epithelial cells)

Get tips on using Live and Dead Cell Assay (Abcam) to perform Live / Dead assay mammalian cells - rabbit bone marrow mesenchymal stem cells

Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Live / Dead assay mammalian cells - mouse bone marrow-derived macrophages

Get tips on using LIVE/DEAD™ Viability/Cytotoxicity Kit, for mammalian cells to perform Live / Dead assay mammalian cells - mouse bone marrow-derived macrophages

Get tips on using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® to perform RNA sequencing Mouse - Bone marrow-derived macrophages (BMDMs)

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.