HT29-MTX-E12
Cat.No.: CSC-C7109J
Species: Homo sapiens (Human)
Source: Intestine; Colon
Morphology: Epithelial
Culture Properties: Adherent
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HT29-MTX-E12 cell line is a well-differentiated, mucus-producing subclone derived from the parent human colorectal adenocarcinoma HT-29 line through long-term adaptation to the chemotherapeutic agent methotrexate (MTX). This selective pressure drives the cells toward a stable goblet cell-like phenotype, characterized by a markedly reduced proliferative rate and a constitutive commitment to differentiation. Unlike the heterogeneous parental line, HT29-MTX-E12 cells consistently express key hallmarks of intestinal goblet cells, most notably the robust production and apical secretion of gel-forming mucins, primarily MUC5AC, forming a functional, viscoelastic mucus layer. This specialized capability establishes it as the gold-standard in vitro model for studying the intestinal mucus barrier, a critical frontline defense system.
It is routinely combined with absorptive enterocyte models (e.g., Caco-2) in transwell co-culture systems to create more physiologically complete intestinal epithelium models. These co-cultures accurately segregate mucus production (HT29-MTX-E12) and tight junction formation (Caco-2), dramatically improving the predictive value for drug permeability, toxicology, and nutraceutical absorption studies.
Protocol For Developing a Mucus-producing Gut-on-a-chip Model from Caco-2 and HT29-MTX-E12 Cells
Organ-on-chip (OoC) technology offers a more relevant gut model in comparison with static in vitro gut models from stable cell lines by replicating the 3D structure, mucus production, fluid dynamics, and mechanical forces, closely simulating in vivo conditions. Here, we present a protocol for developing a mucus-producing gut-on-a-chip (GoC) model from Caco-2 and HT29-MTX-E12 cells. At the end of the experiment, immunofluorescence and confocal microscopy should confirm 3D structure formation, cell polarization, mucus production and tight junction formation in GoC model with relevant cell marker such as VIL1, Muc5AC and ZO1 expression, with Muc5AC expression expected to vary across different cell ratios.
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