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Thrifty, Rapid Intestinal Monolayers (TRIM) Using Caco-2 Epithelial Cells for Oral Drug Delivery Experiments

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Abstract

Purpose

Caco-2 monolayers are the most common model of the intestinal epithelium and are critical to the development of oral drug delivery strategies and gastrointestinal disease treatments. However, current monolayer systems are cost- and/or time-intensive, hampering progress. This study evaluates two separate methods to reduce resource input: FB Essence as a fetal bovine serum (FBS) alternative and a new, 3-day Caco-2 system deemed “thrifty, rapid intestinal monolayers” (TRIM).

Methods

Caco-2 cells were cultured with FB Essence and compared to cells in 10% FBS for proliferation and monolayer formation. TRIM were compared to commonly-used 21-day and Corning® HTS monolayer systems, as well as mouse intestines, for permeability behavior, epithelial gene expression, and tight junction arrangement.

Results

No amount of FB Essence maintained Caco-2 cells beyond 10 passages. In contrast, TRIM compared favorably in permeability and gene expression to intestinal tissues. Furthermore, TRIM cost $109 and required 1.3 h of time per 24-well plate, compared to $164 and 3.7 h for 21-day monolayers, and $340 plus 1.0 h for the HTS system.

Conclusions

TRIM offer a new approach to generating Caco-2 monolayers that resemble the intestinal epithelium. They are anticipated to accelerate the pace of in vitro intestinal experiments while easing financial burden.

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Abbreviations

BIEDE:

Biocoat® Intestinal Epithelium Differentiation Environment

CLDN1:

Claudin 1

DMEM:

Dulbecco’s Modified Eagles Medium

FBE:

FBEssence

FBS:

Fetal bovine serum

FITC-DX4:

4 kDa molecular weight dextran, tagged with fluorescein isothiocyanate

HTS:

Corning® Biocoat® HTS monolayer system

P##:

Passage number ## of Caco-2 cells

PPZ:

1-phenylpiperazine

qPCR:

Quantitative polymerase chain reaction

Rhod123:

Rhodamine 123

SLS:

Sodium lauryl sulfate

TEER:

Transepithelial electrical resistance

TRIM:

Thrifty, rapid intestinal monolayers

ZO-1:

Zonula occludens 1

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Authors and Affiliations

  1. Department of Chemical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, Pennsylvania, 15213–3890, USA

    Nicholas G. Lamson, Rebecca L. Ball, Katherine C. Fein & Kathryn A. Whitehead

  2. Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania, USA

    Kathryn A. Whitehead

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  1. Nicholas G. Lamson
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  2. Rebecca L. Ball
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  4. Kathryn A. Whitehead
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Correspondence to Kathryn A. Whitehead.

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Lamson, N.G., Ball, R.L., Fein, K.C. et al. Thrifty, Rapid Intestinal Monolayers (TRIM) Using Caco-2 Epithelial Cells for Oral Drug Delivery Experiments. Pharm Res 36, 172 (2019). https://doi.org/10.1007/s11095-019-2712-6

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