Establishment of a triple co-culture in vitro cell models to study intestinal absorption of peptide drugs

• Tipo: Artigo em Revista Científica Internacional
• Data: 2013

Autores

Filipa Antunes (Autor) Outra; Fernanda Andrade (Autor) Outra; Francisca Araujo (Autor) Outra; Domingos Ferreira (Autor) FFUP; Bruno Sarmento (Autor) Outra

Revista

Título: European Journal of Pharmaceutics and Biopharmaceutics

Vol. 83

Páginas: 427-435

ISSN: 0939-6411

Editora: Elsevier

Indexação

ISI Web of Knowledge - 114 Citações

Scopus - 162 Citações

Classificação Científica

FOS: Ciências médicas e da saúde > Medicina básica

Outras Informações

ID Authenticus: P-005-25F

DOI: 10.1016/j.ejpb.2012.10.003

Abstract (EN): In vitro cell culture models for studying oral drug absorption during early stages of drug development have become a useful tool in drug discovery and development, with respect to substance throughput and reproducibility. The aim of this study was to establish an in vitro cellular model based on human colon carcinoma Caco-2, mucus-producing HT29, and Rail B cells in order to design a model that more accurately mimics the small intestinal epithelial layer. Normal oriented model was set up by seeding co-cultures of Caco-2 and HT29 cells into Transwell filters and maintained under identical conditions following addition of Raji B to the basolateral chamber. Inverted model was set up seeding Caco-2 and HT29 cells on the basolateral chamber and then transferred in the Transwell device with the epithelial cells facing the basolateral chamber following Raji B addition to the apical compartment. Morphological differences on size and thickness of cell membranes were detected between the models studied by using fluorescence microscopy. On the triple co-culture models, cell membranes were increasing in size and thickness from the Caco-2 to Caco-2/HT29 and Caco-2/Raji B. Also, the nuclei seem to be larger than in the other studied models. Insulin permeation was higher on the triple co-culture model when compared to the Caco-2/HT29 co-culture model. Also, insulin permeation as mediated by nanoparticles and insulin solution permeation was higher on the normal oriented Caco-2/HT29/Raji B model as compared to the inverted model. Overall, our results suggest that Caco-2/HT29/Raji B triple co-culture normal oriented cellular model may be reliable to obtain a more physiological, functional, and reproducible in vitro model of the intestinal barrier to study protein absorption, both in solution and when delivered by nanocarriers.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 9