Stem cell culture media Rat oligodendrocyte precursors (OPCs)

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.

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Found 3 matching solutions for this experiment

Upstream tips
POCs were established from rat neonatal forebrain cells obtained by dissociating cortices in papain
Protocol tips
DMEM supplemented with 10% fetal calf serum (FCS)
Upstream tips
Ten days later, the flasks were shaken for 1 h on an orbital shaker (218 rpm) at 37°C to remove microglia.
Protocol tips
DMEM; ThermoFisher, #11965) containing 20% fetal bovine serum. 
Gibco™DMEM/F-12

Thermo Fisher Scientific

Upstream tips
mNSCs or rNSCs were dissociated to a single cell suspension using Accutase (Gibco). The cells were counted and plated at 4 × 104 cells/cm2 on plates, which were coated with 0.1 mg/mL poly-L-ornithine (Sigma) and then with 10 µg/mL laminin (Sigma), or coated only with 5% Matrigel (BD Biosciences)
Protocol tips
DMEM/F12, Gibco) supplemented with 1× B27 (Gibco), 1× N2 (Gibco), 20 ng/mL FGF2 (Pepro Tech), 20 ng/mL PDGF-AA (Pepro Tech), and 0.4 µM SAG (Enzo Life Sciences) or 200 ng/mL SHH (R&D Systems)
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