Dissecting stromal-epithelial interactions in a 3D in vitro cellularized intestinal model for permeability studies.
Absorption evaluation plays an increasingly important role at the early stage of drug discovery due to its potential to scan the ADME (absorption, distribution, metabolism and excretion) properties of new drug candidates. Therefore, a new three-dimensional (3D) in vitro model replicating the intestinal functioning is herein proposed aiming to dissect the stromal-epithelial interactions and evaluate the permeation of a model drug, insulin. Inspired on the intestinal mucosal architecture, the present model comprises intestinal myofibroblasts (CCD18-Co cells) embedded in Matrigel, onto which epithelial enterocytes (Caco-2 cells) and mucus-producing cells (HT29-MTX cells) were seeded. CCD18-Co myofibroblasts showed to have a central role in the remodeling of the surrounding matrix confirmed by the production of fibronectin. Subsequently, this matrix revealed to be essential to the maintenance of the model architecture by supporting the overlying epithelial cells. In terms of functionality, this model allowed the efficient prediction of insulin permeability in which the presence of mucus, the less tight character between Caco-2 and HT29-MTX epithelial cells and the 3D assembly were critical factors. Concluding, this model constitutes a robust tool in the drug development field with potential to bridge the traditional 2D cell culture models and in vivo animal models.
I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal; FEUP - Faculdade de Engenharia, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal.
Carla
C
Pereira
I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Rua de Jorge Viterbo Ferreira, 4050-313, Porto, Portugal.
Francisca
F
Araújo
I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal.
Cristina C
CC
Barrias
I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal; FEUP - Faculdade de Engenharia, University of Porto, Rua Dr. Roberto Frias, 4200-465, Porto, Portugal; ICBAS - Instituto de Ciências Biomédicas Abel Salazar, University of Porto, Rua de Jorge Viterbo Ferreira, 4050-313, Porto, Portugal.
Pedro L
PL
Granja
I3S - Instituto de Investigação e Inovação em Saúde and INEB - Instituto de Engenharia Biomédica, University of Porto, Rua do Campo Alegre, 823, 4150-180, Porto, Portugal; CESPU, Instituto de Investigação e Formação Avançada em Ciências e Tecnologias da Saúde, Instituto Superior de Ciências da Saúde-Norte, Rua Central de Grandra, 1317, 4585-116, Gandra, Portugal. Electronic address: bruno.sarmento@ineb.up.pt.
Bruno
B
Sarmento
D018938:Caco-2 Cells; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018920:Coculture Techniques / Q000379:methods*; D020895:Enterocytes / Q000166:cytology; D004847:Epithelial Cells / Q000166:cytology*; D005353:Fibronectins / Q000737:chemistry; D019073:HT29 Cells; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007408:Intestinal Absorption; D007413:Intestinal Mucosa / Q000473:pathology; D007422:Intestines / Q000166:cytology*; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D058628:Myofibroblasts / Q000166:cytology; D010539:Permeability; D017154:Stromal Cells / Q000166:cytology*
D005353:Fibronectins; D007328:Insulin
Caco-2 cells; Extracellular matrix (ECM); Fibroblast; HT29-MTX cells; Intestinal mucosa; Triple co-culture