siRNA / miRNA gene silencing Human 501 Mel and SK Mel 28 FANCD2

Get tips on using StemMACS™ AdipoDiff Media, human to perform Stem cell Differentiation media hMSCs differentiation into adipogenic cells

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - human MSCs (mesenchymal stem cells)

Get tips on using PicoGreen® dsDNA Quantitation Reagent and Kit to perform DNA quantification Blood

Get tips on using Human CRISP-3 Antibody to perform Immunohistochemistry Human - CRISP3

Get tips on using Human Myeloperoxidase Quantikine ELISA Kit to perform ELISA Human - MPO

Get tips on using MammoCult™ Human Medium Kit to perform Stem cell culture media hMammospheres

Get tips on using Brilliant Violet 510™ anti-human HLA-DR Antibody to perform Flow cytometry Anti-bodies Human - HLA-DR

Get tips on using Human FGF-10 Antibody to perform Immunohistochemistry Human - FGF-10

A gross majority of classical apoptotic attributes can be quantitatively examined by flow cytometry, the preferred platform for rapid assessment of multiple cellular attributes at a single-cell level. However, sample preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

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.