LIVE IMAGING BY TIME-LAPSE MICROSCOPY CAN MORE CLEARLY EVALUATE THE ANTI-APOPTOTIC STATE OF PRIMARY HEPATOCYTES ISOLATED FROM THE DRS KNOCKOUT MOUSE

概要

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To investigate the anti-apoptotic state of primary hepatocytes isolated from the drs knockout(KO)mice, Fas-mediated apoptosis was observed by flow cytometry and time-lapse microscopy. Treatment with anti-Fas antibody plus actinomycin D or cycloheximide induced over 50% apoptosis for primary hepatocytes based on annexin V staining. At 24 hr later under these apoptotic stimuli, the sub G1 fraction of hepatocytes isolated from the drs KO mice was significantly decreased in comparison to wild C57BL/ 6 mice. In contrast, sequential time-lapse microscopic analysis revealed clearer and larger differences in the apoptotic responses of primary hepatocytes from the drs KO and wild-type mice from 4 hr to 24 hr after apoptotic induction. The time-lapse microscopic analysis clearly indicated the hepatocytes isolated from the drs KO mouse to be more resistant to the apoptotic stimuli than other methods. The time-lapse microscopy is very useful for the sequential observation of viable primary hepatocytes.
秋田大学の論文