title: Design of 3D printed insert for hanging culture of Caco-2 cells.
abstract: A Caco-2 cell culture on Transwell, an alternative testing to animal or human testing used in evaluating drug intestinal permeability, incorrectly estimated the absorption of actively transported drugs due to the low expression of membrane transporters. Similarly, three-dimensional (3D) cultures of Caco-2 cells, which have been recommended to be more physiological relevant, were not superior to the Transwell culture in either accuracy or convenience in drug permeability testing. Using rapid 3D printing prototyping techniques, this study proposed a hanging culture of Caco-2 cells that performed with high accuracy in predicting drug permeability in humans. As found, hanging cultured Caco-2 cells formed a confluent monolayer and maintained high cell viability on the 3D printed insert. Compared with the normal culture on Transwell, the Caco-2 cells on the 3D printed insert presented ∼30-100% higher brush border enzyme activity and ∼2-7 folds higher activity of P-glycoprotein/multidrug resistance-associated protein 2 during 21 days of incubation. For the eight membrane transporter substrates, the predictive curve of the 3D printing culture exhibited better linearity (R(2) = 0.92) to the human oral adsorption than that of the Transwell culture (R(2) = 0.84), indicating better prediction by the 3D printing culture. In this regard, the 3D printed insert for hanging culture could be potentially developed as a convenient and low-cost tool for testing drug oral absorption.
authors.affiliation: Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou, People's Republic of China.
authors.forename: Chong
authors.initials: C
authors.lastname: Shen
authors.forename: Qin
authors.initials: Q
authors.lastname: Meng
authors.forename: Guoliang
authors.initials: G
authors.lastname: Zhang
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D065567:Absorption, Physiological; D018938:Caco-2 Cells; D018929:Cell Culture Techniques / Q000379:methods*; D048430:Cell Shape; D002470:Cell Survival; D006801:Humans; D026901:Membrane Transport Proteins / Q000378:metabolism; D008871:Microvilli / Q000201:enzymology; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism; D004364:Pharmaceutical Preparations / Q000378:metabolism; D066330:Printing, Three-Dimensional; D013997:Time Factors
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D026901:Membrane Transport Proteins; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D004364:Pharmaceutical Preparations