siRNA / RNAi /miRNA transfection Human Cells Jurkat cells

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 Absolutely RNA miRNA Kit to perform RNA isolation / purification Cells - immortalized HT-29

Get tips on using Senescence Cells Histochemical Staining Kit to perform Reporter gene assay β-galactosidase substrates - human MSCs (mesenchymal stem cells)

Get tips on using Stealth siRNA(r)_Plat to perform siRNA / miRNA gene silencing Rat - NPC tPA/Plat

Get tips on using Stealth siRNA(r)_BIRC2 to perform siRNA / miRNA gene silencing Rat - B35 cIAP1/BIRC2

Get tips on using Stealth siRNA(r)_BIRC3 to perform siRNA / miRNA gene silencing Rat - B35 cIAP2/BIRC3

Get tips on using Stealth siRNA(m)_Atg5 to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Atg5

Get tips on using IRF-1 siRNA (m) to perform siRNA / miRNA gene silencing Mouse - B16-F10 IRF1

Get tips on using HIF-1α siRNA (m) to perform siRNA / miRNA gene silencing Mouse - AtT20 Hif-1alpha

Get tips on using siGENOME Rat Epor siRNA to perform siRNA / miRNA gene silencing Rat - H19-7 EpoR