siRNA / RNAi /miRNA transfection Human Cells HESC

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines Neuro2a

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines HeLa

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines BXPC3

Get tips on using pSUPER.retro.neo+gfp vector- Syn G (exon 3) siRNA to perform shRNA gene silencing Mouse - RGC-5 Syn G (Exon 3)

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines SKOV-3

Get tips on using FlexiTube GeneSolution GS27279 for Tnfrsf12a to perform siRNA / miRNA gene silencing Mouse - B16-BL6 FN14/Tnfrsf12a

Get tips on using Lipofectamine® LTX Reagent to perform DNA transfection Mammalian cells - Immortalized cell lines MDA-MB-231

Get tips on using MISSION® esiRNA_esiRNA targeting mouse Lrp5 (esiRNA1) to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Lrp5

Get tips on using MISSION® esiRNA_esiRNA targeting mouse Lrp6 (esiRNA1) to perform siRNA / miRNA gene silencing Mouse - MLO‐Y4 Lrp6

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.