CHARACTERIZATION OF A SPONTANEOUSLY POLARIZING HT-29 CELL LINE, HT-29/cl.f8

Abstract

A cloned cell line that spontaneously polarizes in standard glucose-containing media was derived from a single cell of the adenocarcinoma cell line HT-29. The cloned line, designated HT-29/cl.f8, has remained stable over 2 yr in culture, maintained high transepithelial resistance (300 ohm cm(2) or higher), and correctly sorted influenza virus and vesicular stomatitis virus to apical or basolateral domains, respectively. The newly cloned cells also displayed apical microvilli, tight junctions, and desmosomes, the morphological characteristics of mature epithelia. The cloned HT-29/cl.f8 cells function as epithelial enterocytes as shown by the apical expression of intestinal alkaline phosphatase, the expression of vimentin and cytokeratin, and lack of expression of mucin. We propose that the newly cloned HT-29/cl.f8 cells offer a viable alternative for studies of enterocyte function that will readily yield interpretable data not complicated by cell alterations due to the presence of drugs or chemicals that induce differentiation.

Full-Length, Glycosylated NSP4 Is Localized to Plasma Membrane Caveolae by a Novel Raft Isolation Technique

“…MDCK (American Type Culture Collection, Manassas, VA) and HT29.f8 cells, a spontaneously polarizing cell line derived from the human adenocarcinoma HT29 intestinal cell line (36), were grown in maintenance medium (Dulbecco's modified Eagle's medium [DMEM] with 4.5 g/liter glucose, L-glutamine, and sodium pyruvate; Mediatech, Inc., Herndon, VA) supplemented with 2 mM L-glutamine (BioWhittaker/Cambrex), 1 mM sodium pyruvate (Cambrex), 0.1 mM nonessential amino acids (Mediatech, Inc.), 100 U/liter penicillin, 100 g/liter streptomycin, 0.25 g/liter amphotericin B (10,000 U of penicillin-10,000 U of streptomycin-25 g/liter amphotericin B; Cambrex), 43.9 mM sodium bicarbonate (GIBCO), 5% fetal bovine serum, and 5% Serum Supreme (Cambrex) at 37°C in 5% CO 2 . MDCK cell stocks were maintained in 175-cm 2 flasks and expanded into 500-cm 2 trays or 2-cm 2 multiwell plates (Corning, Inc., Corning, NY) for caveola isolation.…”

Alterations in dysadherin expression and F-actin reorganization: a possible mechanism of hypericin-mediated photodynamic therapy in colon adenocarcinoma cells

“…HT-29 and Caco-2 cells differentiate spontaneously and form typical intestine villus structured colon cancer cells which express specific intestine enzymes (Mitchell and Ball 2004). Previously, HYP-mediated PDT in HT-29 cells was investigated.…”

The Effects of Media on Pharmaceutically Relevant Transporters in the Human HT-29 Adenocarcinoma Cell Line: Does Culture Media Need to be Controlled?

“…21 The HT-29 cell line has also been widely utilized to study nutrient and drug permeation, multidrug resistance, metabolism, and nutrient-induced enterocytic differentiation amongst other pharmaceutically relevant studies. 22 Other studies in the HT-29 cell line include investigating integrin expression, the role of insulin-like growth factors, and mechanisms of lipid metabolism. 22 One of the major differences between this cell line and the more widely utilized Caco-2 cell line is that the HT-29 cells can produce mucin at a relatively high level, which can act as a rate-determining barrier for drug absorption.…”

“…22 Other studies in the HT-29 cell line include investigating integrin expression, the role of insulin-like growth factors, and mechanisms of lipid metabolism. 22 One of the major differences between this cell line and the more widely utilized Caco-2 cell line is that the HT-29 cells can produce mucin at a relatively high level, which can act as a rate-determining barrier for drug absorption. [23][24][25] This is due in part because the parental HT-29 cells are considered to contain a more heterogeneous mixture of cells than the Caco-2 cell model that do not spontaneously differentiate under certain culture conditions.…”