siRNA / miRNA gene silencing Human UCC (urothelial cancer cell lines)

Get tips on using TdT In Situ Apoptosis Detection Kit - Fluorescein to perform TUNEL assay cell type - HeLa cells human cervical cancer

Get tips on using ApopTag Fluorescein in Situ Apoptosis Detection Kit to perform TUNEL assay cell type - PC-3 human prostate cancer

Get tips on using TumorTACS™ In Situ Apoptosis Detection Kit to perform TUNEL assay cell type - PANC-1 human pancriatic cancer

Get tips on using Corning® Hepatocyte Culture Media Kit, 500 mL to perform 3D Cell Culture Media Human bladder cancer organoid

Get tips on using Pierce™ Magnetic ChIP Kit to perform ChIP Human - Fibroblast cell lines

Get tips on using DMEM/F-12, no phenol red to perform 3D Cell Culture Media Human breast cancer MCF-7 cells-Mammospheres

Get tips on using The Premo Autophagy Tandem Sensor RFP-GFP-LC3B Kit to perform Autophagy assay cell type - Human osteosarcoma cancer cells

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.

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.

Get tips on using DMEM/F-12, no phenol red to perform 3D Cell Culture Media Human breast cancer MDA-MB-231 cells-Mammospheres