Peak of DNA fragmentation (apoptotic response) in response to staurosporine treatment (time after treatment)

Range in EL4 (mouse lymphoma cell line) cells 5–6h : in HCE (Human Corneal Epithelial Cells) cells 24h hours
Organism Mammalian tissue culture cell
Reference Sundquist et al. (2006) Timing your apoptosis assays, pp.18-21 link p.19 right column 2nd paragraph
Primary Source [6] Jessel, R. et al. (2002) Kinetics of apoptotic markers in exogeneously induced apoptosis of EL4 cells. J. Cell Mol. Med. 6, 82–92. [7] Härtel S et al., Staurosporine-induced apoptosis in human cornea epithelial cells in vitro. Cytometry A. 2003 Sep55(1):15-23.PubMed ID12003671, 12938184
Method Primary source [6] abstract:"[Researchers] investigated the time-dependence of apoptotic events in EL4 cells by monitoring plasma membrane changes in correlation to DNA fragmentation and cell shrinkage. [They] applied three apoptosis inducers (staurosporine, tubericidine and X-rays) and [they] looked at various markers to follow the early-to-late apoptotic events: phospholipid translocation (identified through annexin V-fluorescein assay and propidium iodide), lipid package (via merocyanine assay), membrane fluidity and anisotropy (via fluorescent measurements), DNA fragmentation by the fluorescence-labeling test and cell size measurements." Primary source [7] abstract:"HCE cells were cultured in the presence of STS [staurosporine] to induce apoptosis. Caspase-3 activity was measured with the fluorogenic substrate z-DEVD-rhodamine 110. [Investigators] determined mitochondrial viability with a 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzenedisulfonate reduction assay, and chromatin degradation with a fluorometric method using 4,6-diamidino-2-phenylindole (DAPI). Membrane translocation of PS and nuclear alterations were assessed by quantitative fluorescence microscopy. Image processing routines were written in interactive data language (IDL)."
Comments P.19 right column 2nd paragraph:"Even with an identical apoptosis-inducing stimulus, timelines for the progression of different cells through apoptosis vary. The time required for an activity to peak in one cell line will likely differ from that for another cell line. For example, DNA fragmentation in response to staurosporine treatment peaked at 5–6 hours in EL4 cells (primary source 6) but at 24 hours in HCE cells (primary source 7). When consulting published protocols to identify apoptosis inducing conditions, do not assume that the effect on your cell line will be the same as that on other cells."
Entered by Uri M
ID 112338