siRNA / miRNA gene silencing Mouse MLO‐Y4

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.

Get tips on using LC3B Mouse Monoclonal Antibody (9H5) to perform Autophagy assay cell type - Ramos

Get tips on using Purified Anti-Mouse TCR beta (H57-597) to perform Flow cytometry Anti-bodies Mouse - TCRbeta

Get tips on using Purified Hamster Anti-Mouse TCR β Chain to perform Flow cytometry Anti-bodies Mouse - TCRbeta

Get tips on using FoxP3 Antibody, anti-mouse, PE, REAfinity™ to perform Flow cytometry Anti-bodies Mouse - FOXP3

Get tips on using Purified anti-mouse CD115 (CSF-1R) Antibody to perform Flow cytometry Anti-bodies Mouse - CD115

Get tips on using Purified NA/LE Hamster Anti-Mouse CD40 to perform Flow cytometry Anti-bodies Mouse - CD40

Get tips on using Mouse Serpin E1/PAI-1 DuoSet ELISA to perform ELISA Mouse - Serpin E1/PAI-1

Get tips on using Mouse C-Reactive Protein/CRP DuoSet ELISA to perform ELISA Mouse - C-Reactive Protein/CRP

Get tips on using Gentra Puregene Mouse Tail Kit (4 g) to perform DNA isolation / purification Tissue - murine tail biopsies