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Induction and detection of apoptosis in BHK 21 C13 cells

Apoptosis is a programmed cell death characterized by specific transformations such as the formation of membrane-enclosed vesicles, cellular shrinkage, chromatin condensation and frag-mentation of the nuclear DNA. Intracellular and plasma membrane structural modifications have been widely recognized as crucial factors involved in cell injury and death. One of the membrane changes in the early/ intermediate stages of apoptosis is the translocation of phosphatidylserine (PS) from the inside to the outside of the cell membrane, with the plasma membrane remaining intact. One of the commonly used methods for PS detection in apoptotic cells is the use of Annexin V, a Ca^2+ dependent phospholipid-binding protein. In this study we have investigated the influence of four different apoptosis stimuli on BHK 21 C13 cells: heat shock, starvation, actinomycin D and dexamethasone. Apoptotic cells were detected by fluorescence microscopy and flow cytometry using a combination of Annexin V labelled with FITC and propidium iodide (PI). We have found that BHK 21 C13 cells undergo apoptosis under nonphysiological conditions: heat shock and starvation. Actinomycin D (5 microg/mL) induced apoptosis in BHK 21 C13 cells, whereas dexamethasone treatment did not.

apoptosis, BHK 21 C13 cell line, flow cytometry, phosphatidylserine (PS), Annexin V

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