title: Hollow fiber culture accelerates differentiation of Caco-2 cells.
abstract: Caco-2 cells usually require 21 days of culture for developing sufficient differentiation in traditional two-dimensional Transwell culture, deviating far away from the quick differentiation of enterocytes in vivo. The recently proposed three-dimensional cultures of Caco-2 cells, though imitating the villi/crypt-like microstructure of intestinal epithelium, showed no effect on accelerating the differentiation of Caco-2 cells. In this study, a novel culture of Caco-2 cells on hollow fiber bioreactor was applied to morphologically mimic the human small intestine lumen for accelerating the expression of intestine functions. The porous hollow fibers of polyethersulfone (PES), a suitable membrane material for Caco-2 cell culture, successfully promoted cells to form confluent monolayer on the inner surface. The differentiated functions of Caco-2 cells, represented by alkaline phosphatase, γ-glutamyltransferase, and P-glycoprotein activity, were greatly higher in a 10-day hollow fiber culture than in a 21-day Transwell culture. Moreover, the Caco-2 cells on PES hollow fibers expressed higher F-actin and zonula occludens-1 protein than those on Transwell culture, indicative of an increased mechanical stress in Caco-2 cells on PES hollow fibers. The accelerated differentiation of Caco-2 cells on PES hollow fibers was unassociated with membrane chemical composition and surface roughness, but could be stimulated by hollow fiber configuration, since PES flat membranes with either rough or smooth surface failed to enhance the differentiation of Caco-2. Therefore, the accelerated expression of Caco-2 cell function on hollow fiber culture might show great values in simulation of the tissue microenvironment in vivo and guide the construction of intestinal tissue engineering apparatus.
authors.affiliation: Department of Chemical and Biological Engineering, Zhejiang University, 38 Zheda Road, Hangzhou, Zhejiang, 310027, People's Republic of China.
authors.forename: Xudong
authors.initials: X
authors.lastname: Deng
authors.forename: Guoliang
authors.initials: G
authors.lastname: Zhang
authors.forename: Chong
authors.initials: C
authors.lastname: Shen
authors.forename: Jian
authors.initials: J
authors.lastname: Yin
authors.forename: Qin
authors.initials: Q
authors.lastname: Meng
mesh_terms: D001483:Base Sequence; D019149:Bioreactors; D018938:Caco-2 Cells; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D017931:DNA Primers; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers