Abstract
The aim of this interdisciplinary project is to establish slice culture preparations from rodents and humans as a new model system for studying effects of X-rays and heavy ions within normal and tumor tissues. The advantage of such slice cultures relies on the conservation of an organotypic environment, the easy treatment and observation by live-imaging microscopy, and the independence from genetic immortalization strategies used to generate cell lines. Rat brains as well as human tumors were cut into 300-μm-thick sections and cultivated in an incubator in a humidified atmosphere at 37°C. This is realized by a membrane-based culture system with a liquid–air interface. With this system, it is possible to keep rodent slices viable for several months. Human brain tumor slices remained vital for at least 21 days. Slices were irradiated with X-rays at the radiation facility of the University Hospital in Frankfurt/Main at doses up to 40 Gy. Heavy ion irradiations were performed at GSI (Darmstadt) with different ions, energies, and doses. The irradiated slices were analyzed by 3D-confocal microscopy following immunostaining for DNA damage, microglia, and proliferation markers. The phosphorylated histone γH2AX proved to be suitable for the detection of ion traversals in this system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aktas O, Smorodchenko A, Brocke S, Infante-Duarte C, Schulze Topphoff U, Vogt J, Prozorovski T, Meier S, Osmanova V, Pohl E, Bechmann I, Nitsch R, Zipp F (2005) Neuronal damage in autoimmune neuroinflammation mediated by the death ligand TRAIL. Neuron 46:421–432
Belyakov OV, Mitchell SA, Parikh D, Randers-Pehrson G, Marino SA, Amundson SA, Geard CR, Brenner DJ (2005) Biological effects in unirradiated human tissue induced by radiation damage up to 1 mm away. Proc Natl Acad Sci USA 4(102):14203–14208
Dehghani F, Hischebeth GT, Wirjatijasa F, Kohl A, Korf HW, Hailer NP (2003) The immunosuppressant mycophenolate mofetil attenuates neuronal damage after excitotoxic injury in hippocampal slice cultures. Eur J Neurosci 18:1061–1072
Dowsing T, Kendall MJ (2007) The Northwick Park tragedy—protecting healthy volunteers in future first-in-man trials. J Clin Pharm Ther 32:203–207
Eyüpoglu IY, Bechmann I, Nitsch R (2003) Modification of microglia function protects from lesion-induced neuronal alterations and promotes sprouting in the hippocampus. FASEB J 17:1110–1111
Hailer NP, Jarhult JD, Nitsch R (1996) Resting microglial cells in vitro: analysis of morphology and adhesion molecule expression in organotypic hippocampal slice cultures. Glia 18:319–331
Hailer NP, Grampp A, Nitsch R (1999) Proliferation of microglia and astrocytes in the dentate gyrus following entorhinal cortex lesion: a quantitative bromodeoxyuridine-labelling study. Eur J Neurosci 11:3359–3364
Hailer NP, Wirjatijasa F, Roser N, Hischebeth GT, Korf HW, Dehghani F (2001) Astrocytic factors protect neuronal integrity and reduce microglial activation in an in vitro model of N-methyl-D-aspartate-induced excitotoxic injury in organotypic hippocampal slice cultures. Eur J Neurosci 14:315–326
Jakob B, Scholz M, Taucher-Scholz G (2003) Biological imaging of heavy charged-particle tracks. Radiat Res 159:676–684
Kenter MJ, Cohen AF (2006) Establishing risk of human experimentation with drugs: lessons from TGN1412. Lancet 368:1387–1391
Kluge A, Hailer NP, Horvath TL, Bechmann I, Nitsch R (1998) Tracing of the entorhinal-hippocampal pathway in vitro. Hippocampus 8:57–68
Kraft G, Daues HW, Fischer B, Kopf U, Liebold HP, Quis DD, Stelzer H, Kiefer J, Schöpfer F, Schneider E, Weber KJ, Wulf H, Dertinger H (1980) Irradiation chamber and sample changer for biological samples. Nucl Instr Meth 168:175–179
Kreutz S, Koch M, Ghadban C, Korf HW, Dehghani F (2007) Cannabinoids and neuronal damage: differential effects of THC, AEA and 2-AG on activated microglial cells and degenerating neurons in excitotoxically lesioned rat organotypic hippocampal slice cultures. Exp Neurol 203:246–257
Kreutz S, Koch M, Böttger C, Ghadban C, Korf HW, Dehghani F (2009) 2-Arachidonoylglycerol elicits neuroprotective effects on excitotoxically lesioned dentate gyrus granule cells via abnormal-cannabidiol-sensitive receptors on microglial cells. Glia 57:286–294
Kreutzberg GW (1996) Microglia: a sensor for pathological events in the CNS. Trends Neurosci 19:312–318
Nitsch R, Bechmann I, Deisz RA, Haas D, Lehmann TN, Wendling U, Zipp F (2000) Human brain-cell death induced by tumour-necrosis-factor-related apoptosis-inducing ligand (TRAIL). Lancet 356:827–828
Nitsch R, Pohl EE, Smorodchenko A, Infante-Duarte C, Aktas O, Zipp F (2004) Direct impact of T cells on neurons revealed by two-photon microscopy in living brain tissue. J Neurosci 24:2458–2464
Roig AI, Hight SK, Shay JW (2009) Two- and three-dimensional models for risk assessment of radiation-enhanced colorectal tumorigenesis. Radiat Res 171:33–40
Schaefer L, Babelova A, Kiss E, Hausser HJ, Baliova M, Krzyzankova M, Marsche G, Young MF, Mihalik D, Götte M, Malle E, Schaefer RM, Gröne HJ (2005) The matrix component biglycan is proinflammatory and signals through Toll-like receptors 4 and 2 in macrophages. J Clin Invest 115:2223–2233
Sowa MB, Chrisler WB, Zens KD, Ashjian EJ, Opresko LK (2010). Three-dimensional culture conditions lead to decreased radiation induced cytotoxicity in human mammary epithelial cells. Mutat Res. doi:10.1016/j.mrfmmm.2010.03.004
Su Y, Meador JA, Geard CR, Balajee AS (2009) Analysis of ionizing radiation-induced DNA damage and repair in three-dimensional human skin model system. Exp Dermatol. doi:10.1111/j.1600-0625.2009.00945.x
Ullrich O, Diestel A, Eyüpoglu IY, Nitsch R (2001) Regulation of microglial expression of integrins by poly(ADP-ribose) polymerase-1. Nat Cell Biol 3:1035–1042
Walczak H, Miller RE, Ariail K, Gliniak B, Griffith TS, Kubin M, Chin W, Jones J, Woodward A, Le T, Smith C, Smolak P, Goodwin RG, Rauch CT, Schuh JC, Lynch DH (1999) Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 5:157–163
Acknowledgments
This project has been funded by the Kassel-Stiftung, the Messer-Stiftung, the Beilstein-Stiftung and the Dr. Senckenbergische-Stiftung. We are truly grateful for the start-up funding and the since then continuous support of these Frankfurt foundations. Aspects of our work with slice cultures are now financed by the Bundesministerium für Bildung und Forschung (BMBF to I.B.) and the European Space Association (ESA to I.B. and G. T.-S.).
Author information
Authors and Affiliations
Corresponding author
Additional information
This manuscript is based on a contribution given at the “Heavy Ions in Therapy and Space Symposium 2009,” July 6–10, 2009, Cologne (Germany).
F. Merz and M. Müller contributed equally to the manuscript.
Rights and permissions
About this article
Cite this article
Merz, F., Müller, M., Taucher-Scholz, G. et al. Tissue slice cultures from humans or rodents: a new tool to evaluate biological effects of heavy ions. Radiat Environ Biophys 49, 457–462 (2010). https://doi.org/10.1007/s00411-010-0293-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00411-010-0293-1