Site Directed Mutagenesis (SDM) Human Point mutation SH-SY5Y

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 RNeasy Mini Kit to perform RNA isolation / purification Tissue - Human Seminal vesicles

Get tips on using ChIP Kit Magnetic - One Step (ab156907) to perform ChIP Human - MIA PaCa-2

Get tips on using ChIP Kit Magnetic - One Step (ab156907) to perform ChIP Human - MDA-MB-231

Get tips on using siRNA Wee1 to perform siRNA / miRNA gene silencing Human - SKOV-3 Wee-1

Get tips on using siRNA RBM3 to perform siRNA / miRNA gene silencing Human - MIA PaCa-2 RBM3

Get tips on using siRNA RIP3 to perform siRNA / miRNA gene silencing Human - MIA PaCa-2 RIP3

Get tips on using siRNA Wee1 to perform siRNA / miRNA gene silencing Human - MDA-MB-231 Wee1

Get tips on using siRNA FOXM1 to perform siRNA / miRNA gene silencing Human - MDA-MB-231 FOXM1

Get tips on using Hs_PTEN_6 FlexiTube siRNA to perform siRNA / miRNA gene silencing Human - HCT-116 PTEN1