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United States Department of Agriculture

Agricultural Research Service

Title: Support Membrane Porosity Influences the Morphological Uniformity of Caco-2cells in Culture

Authors
item Briske Anderson, Mary
item Newman Jr, Samuel
item Reeves, Phillip

Submitted to: Federation of American Societies for Experimental Biology Conference
Publication Type: Abstract Only
Publication Acceptance Date: April 18, 1998
Publication Date: N/A

Technical Abstract: The CACO-2 cell is a popular model for intestinal absorption of trace elements. CACO-2 cells grown on various support surfaces develop topological departures from the optimal monolayer. Hollow domes and solid multicellular protrusions occur on solid supports. On porous membranes used for transport studies the solid protrusions of varied size and shape contribute an indeterminate amount of surface area. This complicates measurements of rates of transport. The purpose of this study was to determine the effect different porosities of support membranes have on the development of these structures. CACO-2 cells were seeded onto collagen treated porous membranes made of polyethyleneterephthalate (PET) with pore sizes of 0.45, 1, and 3 :m or polycarbonate, pore size 0.45 :m. The 0.45 :m membranes were available in two porosities, low (1.6 x 106 pores/cm2) and high (1.0 x 108 pores/cm2) density. Duplicate cultures were embedded in epoxy resin at growth day 21. Light microscopy of sectioned CACO-2 cultures showed that domes do not form on porous supports. Multicellular protrusions, however, do occur on porous membranes. Upon comparing cultures on membranes with pores of different sizes and density distributions, there was an apparent decrease in both the number and size of these protrusions with an increase in pore density; this was not affected by pore size. While protrusions were not completely eliminated on 0.45 um high porosity membranes, we believe that the more uniform cultures these membranes provide are better for transport studies.

Last Modified: 11/27/2014
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