RNA isolation / purification Cells primary

Get tips on using SV 96 Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized T84

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.

ROS has a very short half-lives in biological environment as they are influenced by exposure to ambient oxygen. As it is highly reactive and hard to measure care should be taken to ensure the stability of the sample during isolation, preparation, storage, and analysis.

Get tips on using RNAqueous®-Micro Total RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized T47D

Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized SK-BR-3

Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized MDA-MB-231

Get tips on using SV Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized U-2 OS

Get tips on using SV 96 Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized PK-1

Get tips on using SV 96 Total RNA Isolation System to perform RNA isolation / purification Cells - immortalized Calu-3

Get tips on using miRCURY RNA Isolation Kit to perform RNA isolation / purification Cells - immortalized SKBR3, MDA-MB231 and MCF7