- Research Article
- Open Access
DMSO modulates the pathway of apoptosis triggering
Cellular & Molecular Biology Letters volume 16, pages328–341 (2011)
We demonstrate here that distribution of caspase-9 influences the pathway of apoptosis triggering, since caspase-9 is activated efficiently only when it is distributed solely in the cytosol. Caspase-9 moves to the nuclei in a response to cell stress during isolation of primary hepatocytes; this is called preapoptotic cell stress response. The dimethyl sulfoxide (DMSO) treatment cannot prevent the migration of caspase-9 into the nuclei when it is added to primary hepatocytes immediately after isolation; however, it can trigger redistribution of caspase-9 from the nuclei into the cytosol when added 1 day post-isolation. This redistribution is temporary, since caspase-9 returns to the nuclei within 48 hours of DMSO treatment. Thereafter, some caspase-9 is retained in the nuclei of DMSO-treated hepatocytes for longer than in the nuclei of untreated hepatocytes. By measuring caspase activities, we demonstrate that the addition of DMSO to cell culture medium can temporarily normalize the susceptibility of hepatocytes for apoptosis triggering through the intrinsic pathway. DMSO contributes also to the prolonged pathway inactivation, i.e., by extending preapoptotic cell stress response. We propose that DMSO extends the survival of primary hepatocytes by modulating preapoptotic cell stress response, which could be exploited for extending the lifespan of other primary cell cultures.
Chinese hamster ovary
fetal calf serum
Puppi, J. and Dhawan, A. Human hepatocyte transplantation overview. Methods. Mol. Biol. 481 (2009) 1–16.
Strom, S.C., Fisher, R.A., Thompson, M.T., Sanyal, A.J., Cole, P.E., Ham, J.M. and Posner, M.P. Hepatocyte transplantation as a bridge to orthotopic liver transplantation in terminal liver failure. Transplantation 63 (1997) 559–569.
Bilir, B.M., Guinette, D., Karrer, F., Kumpe, D.A., Krysl, J., Stephens, J., McGavran, L., Ostrowska, A. and Durham, J. Hepatocyte transplantation in acute liver failure. Liver Transpl. 6 (2000) 32–40.
Schneider, A., Attaran, M., Meier, P.N., Strassburg, C., Manns, M.P., Ott, M., Barthold, M., Arseniev, L., Becker, T. and Panning, B. Hepatocyte transplantation in an acute liver failure due to mushroom poisoning. Transplantation 82 (2006) 1115–1116.
Horslen, S.P., McCowan, T.C., Goertzen, T.C., Warkentin, P.I., Cai, H.B., Strom, S.C. and Fox, I.J. Isolated hepatocyte transplantation in an infant with a severe urea cycle disorder. Pediatrics 111 (2003) 1262–1267.
Stephenne, X., Najimi, M., Sibille, C., Nassogne, M.C., Smets, F. and Sokal, E.M. Sustained engraftment and tissue enzyme activity after liver cell transplantation for argininosuccinate lyase deficiency. Gastroenterology 130 (2006) 1317–1323.
Muraca, M., Gerunda, G., Neri, D., Vilei, M.T., Granato, A., Feltracco, P., Meroni, M., Giron, G. and Burlina, A.B. Hepatocyte transplantation as a treatment for glycogen storage disease type 1a. Lancet 359 (2002) 317–318.
Fitzpatrick, E., Mitry, R.R. and Dhawan, A. Human hepatocyte transplantation: state of the art. J. Intern. Med. 266 (2009) 339–357.
Fox, I.J., Chowdhury, J.R., Kaufman, S.S., Goertzen, T.C., Chowdhury, N.R., Warkentin, P.I., Dorko, K., Sauter, B.V. and Strom, S.C. Treatment of the Crigler-Najjar syndrome type I with hepatocyte transplantation. N. Engl. J. Med. 338 (1998) 1422–1426.
Ambrosino, G., Varotto, S., Strom, S.C., Guariso, G., Franchin, E., Miotto, D., Caenazzo, L., Basso, S., Carraro, P., Valente, M.L., D’Amico, D., Zancan, L. and D’Antiga, L. Isolated hepatocyte transplantation for Crigler-Najjar syndrome type 1. Cell. Transplant. 14 (2005) 151–157.
Gomez-Lechon, M.J., Donato, M.T., Castell, J.V. and Jover, R. Human hepatocytes as a tool for studying toxicity and drug metabolism. Curr. Drug. Metab. 4 (2003) 292–312.
Liang, J. F. and Akaike, T. Dimethyl sulfoxide induces multilayer aggregates and prolongs survival of primary cultured hepatocytes. Biotechnol. Tech. 11 (1997) 869–872.
Oberhammer F.A., Pavelka M., Sharma S., Tiefenbacher R., Purchio A.F., Bursch W. and Schulte-Hermann R. Induction of apoptosis in cultured hepatocytes and in regressing liver by transforming growth factor beta 1. Proc. Natl. Acad. Sci. USA 89 (1992) 5408–5412.
Salvesen, G.S. and Dixit, V.M. Caspases: intracellular signaling by proteolysis. Cell 91 (1997) 443–446.
Tafani, M., Minchenko, D.A., Serroni, A. and Farber, J.L. Induction of the mitochondrial permeability transition mediates the killing of HeLa cells. Cancer Res. 61 (2001) 2459–2466.
Zhang, X.D., Gillespie, S.K. and Hersey P. Staurosporine induces apoptosis of melanoma by both caspase-dependent and — independent apoptotic pathways. Mol. Cancer Ther. 3 (2004) 187–197.
Nipič, D., Pirc, A., Banič, B., Šuput, D. and Milisav, I. Preapoptotic cell stress response of primary hepatocytes. Hepatology 51 (2010) 2140–2151.
Tuschl, G. and Mueller, S.O. Effects of cell culture conditions on primary rat hepatocytes-cell morphology and differential gene expression. Toxicology 218 (2006) 205–215.
Rowley, S.D., Bensinger, W.I., Gooley, T.A. and Buckner, C.D. Effect of cell concentration on bone marrow and peripheral blood stem cell cryopreservation. Blood 83 (1994) 2731–2736.
Sosef, M.N., Baust, J.M., Sugimachi, K., Fowler, A., Tompkins, R.G. and Toner, M. Cryopreservation of isolated primary rat hepatocytes: enhanced survival and long-term hepatospecific function. Ann. Surg. 241 (2005) 125–133.
Fiore, M. and Degrassi, F. Dimethyl sulfoxide restores contact inhibitioninduced growth arrest and inhibits cell density-dependent apoptosis in hamster cells. Exp. Cell. Res. 251 (1999) 102–110.
Trubiani, O., Ciancarelli, M., Rapino, M. and Di Primio, R. Dimethyl sulfoxide induces programmed cell death and reversible G1 arrest in the cell cycle of human lymphoid pre-T cell line. Immunol. Lett. 50 (1996) 51–57.
Galbraith, R.A., Sassa, S. and Kappas, A. Induction of haem synthesis in Hep G2 human hepatoma cells by dimethyl sulphoxide. A transcriptionally activated event. Biochem. J. 237 (1986) 597–600.
Isom, H.C., Secott, T., Georgoff, I., Woodworth, C. and Mummaw, J. Maintenance of differentiated rat hepatocytes in primary culture. Proc. Natl. Acad. Sci. USA 82 (1985) 3252–3256.
Bour, E.S., Ward, L.K., Cornman, G.A. and Isom, H.C. Tumor necrosis factor-alpha-induced apoptosis in hepatocytes in long-term culture. Am. J. Pathol. 148 (1996) 485–495.
Isom, I., Georgoff, I., Salditt-Georgieff, M. and Darnell Jr., J.E., Persistence of liver-specific messenger RNA in cultured hepatocytes: different regulatory events for different genes. J. Cell. Biol. 105 (1987) 2877–2885.
Milisav, I., Nipič, D. and Šuput, D. The riddle of mitochondrial caspase-3 from liver. Apoptosis 14 (2009) 1070–1075.
Berridge, M.V., Herst, P.M. and Tan, A.S. Tetrazolium dyes as tools in cell biology: new insights into their cellular reduction. Biotechnol. Annu. Rev. 11 (2005) 127–152.
These authors contributed equally to this work