The achievement of the long-term objective of animal replacement in systemic toxicity testing will ultimately depend on the successful establishment and validation of alternative in vitro cellular systems. In modern toxicology, there is an urgent need for efficient and cost-effective in vitro tools to predict pharmacokinetics and toxic reactions of new compounds at early stage of drug discovery.
Biotransformation occurs mainly in the liver, after which hepatocyte-end products are transported to other organs, such as the brain. We have been working on new tools, with higher predictive power in terms of xenobiotic biotransformation than the currently available ones, using primary cultures of hepatocytes (rat and human origin) and differentiating human embryonic stem cells (hESC) into functional hepatocyte-like cells.
- Stock K, Estrada MF, Vidic S, Gjerde K, Rudisch A, Santo VE, Barbier M, Blom S, Arundkar SC, Selvam I, Osswald A, Stein Y, Gruenewald S, Brito C, van Weerden W, Rotter V, Boghaert E, Oren M, Sommergruber W, Chong Y, de Hoogt R, Graeser R. 2016. Capturing tumor complexity in vitro: Comparative analysis of 2D and 3D tumor models for drug discovery. Scientific Reports. 6, 28951
- Santo VE, Rebelo SP, Estrada MF, Alves PM, Boghaert E, Brito C. 2016. Drug screening in 3D in vitro tumor models: overcoming current pitfalls of efficacy read-outs. Biotechnology Journal.
- Terrasso AP, Silva AC, Filipe A, Pedroso P, Ferreira AL, Alves PM, Brito C. 2016. Human neuron-astrocyte 3D co-culture-based assay for evaluation of neuroprotective compounds. J Pharmacol Toxicol Methods. 83:72-79.
- Santo VE, Estrada MF, Rebelo SP, Abreu S, Silva I, Pinto C, Veloso SC, Serra AT, Boghaert E, Alves PM, Brito, C. 2016. Adaptable stirred-tank culture strategies for large scale production of multicellular spheroid-based tumor cell models. J Biotechnol. 221 118-29.
- Estrada MF, Rebelo SP, Davies EJ, Pinto MT, Pereira H, Santo VE, Matthew J. Smalley, Barry ST, Gualda EJ, Alves PM, Anderson E, Brito C. 2016. Modelling the tumor microenvironment in long-term microencapsulated 3D cocultures recapitulates phenotypic features of disease progression. Biomaterials.
- Rebelo SP, Costa R, Silva MM, Marcelino P, Brito C, Alves PM. 2015. Three-dimensional co-culture of human hepatocytes and mesenchymal stem cells: improved functionality in long-term bioreactor cultures. J Tissue Eng Regen Med.
- Rebelo SP, Costa R, Sousa MF, Brito C, Alves PM. 2015. Establishing Liver Bioreactors for In Vitro Research. Methods Mol Biol. 1250:189-202.
- Materne EM, Ramme AP, Terrasso AP, Serra M, Alves PM, Brito C, Sakharov DA, Tonevitsky AG, Lauster R, Marx U. 2015. A multi-organ chip co-culture of neurospheres and liver equivalents for long-term substance testing. J Biotechnol. Feb 9. pii: S0168-1656(15)00045-0.
- Suter-Dick L, Alves PM, Blaauboer BJ, Bremm KD, Brito C, Coecke S, Flick B, Fowler P, Hescheler J, Ingelman-Sundberg M, Jennings P, Kelm JM, Manou I, Mistry P, Moretto A, Roth A, Stedman D, van de Water B, Beilmann M. 2015. Stem Cell-Derived Systems in Toxicology Assessment. Stem Cells Dev. Mar 31. [Epub ahead of print]
- Terrasso AP, Pinto C, Serra M, Filipe A, Almeida S, Ferreira AL, Pedroso P, Brito C, Alves PM. Novel scalable 3D cell based model for in vitro neurotoxicity testing: Combining human differentiated neurospheres with gene expression and functional endpoints. J Biotechnol. 2015 Jan 5. pii: S0168-1656(14)01047-5.
- Hickman J, Graeser R, De Hoogt R, Vidic S, Brito C, Gutekunst M, Van der Kuip H. 2014. Capturing tumor complexity in vitro/ex vivo: three dimensional models of cancer for pharmacology and cancer cell biology. Biotechnology Journal. Biotechnology Journal, 9: 1115-28.
- Rebelo SP, Costa R, Estrada M, Shevchenko V, Brito C, Alves PM. 2014. HepaRG microencapsulated spheroids in DMSO‑free culture: novel culturing approaches for enhanced xenobiotic and biosynthetic metabolism. Archives of Toxicology. Published online 09 August 2014.
- Tostões R, Leite SB, Serra M, Jensen J, Björquist P, Carrondo MJ, Brito C, Alves PM, 2012. Human liver cell spheroids in extended perfusion bioreactor culture for repeated dose drug testing. Hepatology, 55(4): 1227-36.
- Serra M, Brito C, Correia C, Alves PM, 2012. Process engineering of human pluripotent stem cells for clinical applications. Trends Biotechnol, 30(6):350-9.
- Brito C, Simão D, Costa I, Malpique R, Pereira CI, Fernandes P, Serra M, Schwarz SC, Schwarz J, Kremer EJ, Alves PM, 2012. Generation and genetic modification of 3D cultures of human dopaminergic neurons derived from neural progenitor cells. Methods, 56(3):452-60.
- Leite SB, Wilk-Zasadna I, Zaldivar Comenges JM, Airola E, Reis-Fernandes MA, Mennecozzi M, Guguen-Guillouzo C, Chesne C, Gouillou C, Alves PM, Coecke S, 2012. 3D HepaRG model as an attractive tool for toxicity testing. Toxicol Sci 130 (1): 106-116.
- Tostões RM, Leite SB, Miranda JP, Sousa MF, Wang DIC, Carrondo MJ, Alves PM, 2011. Perfusion of 3D encapsulated hepatocytes-a synergistic effect enhancing long-term functionality in bioreactors. Biotechnol Bioeng, 108: 41-49.
- Leite SB, Teixeira AP, Miranda JP, Tostoes RM, Clemente JJ, Sousa MF, Carrondo MJ, Alves PM, 2011. Merging bioreactor technology with 3D hepatocyte-fibroblast culturing approaches: improved in vitro models for Toxicological applications. Toxicol in Vitro, 25(4):825-32.
- Miranda JP, Rodrigues A, Tostões RM, Leite SB, Zimmerman H, Carrondo MJ, Alves PM, 2010. Extending hepatocyte functionality for drug-testing applications using high-viscosity alginate-encapsulated three-dimensional cultures in bioreactors. Tissue Eng Part C Methods, 16(6):1223-32.
- Miranda JP, Leite SB, Muller-Vieira U, Rodrigues A, Carrondo MJ, Alves PM, 2009. Toward extended functional hepatocyte in vitro culture. Tissue Eng Part C Methods, 15:1-11.