Get tips on using ToxCount™ Cell Viability Assay to perform Live / Dead assay mammalian cells - glioblastoma stem cells
Get tips on using Cellular ROS/Superoxide Detection Assay Kit to perform ROS assay cell type - Raw 264.7
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.
DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.
Get tips on using In Vitro Toxicology Assay Kit, Lactic Dehydrogenase based to perform Cell cytotoxicity / Proliferation assay cell type - THP-1
Get tips on using ROS-Glo™ H2O2 Assay to perform ROS assay cell type - MDA-MB-231
Get tips on using PE Annexin V Apoptosis Detection Kit I to perform Apoptosis assay cell type - T-cells Mouse (CD4+ and CD8+)
Cell cycle can be challenging due to difference introduced by sample handling, timing, and difference within the sample. Downstream instriuments to analyse cell cycle (Multicolor flow cytometry and multicolor imaging) can answer these challenges. Relevant markers can be combined with cell phenotyping markers to look at events within subpopulations of cells.
Get tips on using CytoSelect™ 24-Well Cell Migration and Invasion Assay Combo Kit, 8 µm to perform Cell migration / Invasion cell type - BRO
Get tips on using CytoSelect™ 24-Well Cell Migration and Invasion Assay Combo Kit, 8 µm to perform Cell migration / Invasion cell type - LNCaP
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