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. 2009 May;4(3):397-406.
doi: 10.2217/rme.09.4.

Co-culture of intestinal epithelial and stromal cells in 3D collagen-based environments

M E Viney  1 , A J Bullock, M J Day, S MacNeil
Affiliations

Co-culture of intestinal epithelial and stromal cells in 3D collagen-based environments

M E Viney et al. Regen Med. 2009 May.

Abstract

Aim: To investigate the co-culture of established intestinal epithelial cell lines and stromal cells in a series of collagen-based environments for production of tissue-engineered intestinal epithelium for in vitro investigations.

Materials & methods: Intestinal epithelial cells were co-cultured with fibroblasts on a range of supporting collagen matrices including commercially available Promogran and on collagen-based gels.

Results: Epithelial growth was achieved with one combination of vimentin-expressing stromal and cytokeratin-expressing intestinal epithelial cells grown on collagen gels supplemented with Matrigel, and held at an air-liquid interface.

Conclusions: Collagen-based gels can support the co-culture of intestinal epithelial and stromal cells resulting in the growth of an epithelium that has some morphological similarity to normal intestinal tissue.

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Figures

Figure 1
Figure 1
The morphology of (a) Rat-2, (b) IEC 6, (c) IPI-21 and (d) CRL-2012 when grown in submerged monoculture. Bar = 10μm.
Figure 2
Figure 2
The morphology of 2D co-cultures of Rat-2 with (a) IEC 6, (b) IPI-21 and (c) CRL-2102 at different epithelial cell : Rat-2 ratios, immunolabelled for cytokeratin expression. Bar = 10μm.
Figure 3
Figure 3
The morphology of seven-day-old 3D mono- and Rat-2-co-cultures of (a) IEC 6, (b) IPI-21 and (c) CRL-2102 on Promogran held at the air-liquid interface and (d) immunohistochemical labeling for vimentin and cytokeratin of (c). Bar = 10μm.
Figure 4
Figure 4
The morphology of 3D co-cultures on collagen gels of Rat-2 with (a) IEC 6, (b) IPI-21 and (c) CRL-2102. Bar = 10μm.
Figure 5
Figure 5
The morphology of 3D co-cultures on collagen-Matrigel gels of Rat-2 with (a) IEC 6, (b) IPI-21, and CRL-2102 at (c) and (d) 15 and (e) 20 days. Bar = 10μm.
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