siRNA / miRNA gene silencing Human Primary Endometrial Stromal Cells

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human blood endothelial cell

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 Cells - primary human fibroblast-like synoviocytes

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - primary human islets of langerhans

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human dermal fibroblasts

Get tips on using AllPrep RNA/Protein Kit to perform RNA isolation / purification Cells - primary human epidermal keratinocytes

Get tips on using TRI Reagent® Sigma to perform RNA isolation / purification Cells - primary human cardiac fibroblasts

Get tips on using RNeasy Plus Mini Kit to perform RNA isolation / purification Cells - primary human CD14+ monocytes

Get tips on using PureLink™ RNA Mini Kit to perform RNA isolation / purification Cells - primary human chondrocytes