Gaggioli 2007 suppl - S U P P L E M E N TA R Y I N F O R M...

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© 200 7 Nature Publishing Group SUPPLEMENTARY INFORMATION WWW.NATURE.COM/NATURECELLBIOLOGY 1 SCC cells collagen I + laminin matrix supporting mesh media fibroblast direction of invasion 0 0.1 0.2 0.3 0.4 0.5 0.6 Invasion index in gel - cond. media fibroblasts A D C B f scc scc SCC invasion index .5 5.0 10.0 Fibroblasts (*10 5 ) -2 0 0.2 0.4 0.6 0.8 1 5.0 tumour A tumour B ** ** ** A431 A431 + fibroblasts HaCAT + fibroblasts E HT1080 + fibroblasts HT1080 Figure S1 Organotypic culture model for invasion of carcinoma cells with epithelial characteristics. A) Schematic representation of organotypic culture system for studying squamous cell carcinoma invasion. B) Quantification of SCC12 invasion in an organotypic system in the absence, presence of varying numbers of stromal fibroblasts isolated from two different Squamous Cell Carcinomas (SCC’s): Tumour A – head and neck SCC, Tumour B – vulval SCC (average of two experiments - * indicates p<0.05, ** indicates p<0.01). C) Representative images of invasion of A431 (SCC cells), HaCAT (untransformed keratinocytes) and HT1080 (fibrosarcoma cells) in the presence or absence of stromal fibroblasts. D) E-cadherin (red) and DAPI (green) staining of collectively invading SCC cells, ‘f’ indicates fibroblasts ‘scc’ indicates squamous cell carcinoma cells. E) Quantification of SCC12 invasion in an organotypic system in the absence, presence of stromal fibroblasts or media conditioned by stromal fibroblasts (average of two experiments).
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© 200 7 Nature Publishing Group SUPPLEMENTARY INFORMATION 2 WWW.NATURE.COM/NATURECELLBIOLOGY Figure S2 Spatial relationship of stromal fibroblasts and SCC cells during collective invasion. each square represents an individual cell (red shades represent stromal fibroblasts, green shades represent SCC cells), the chain length is represented by the Y axis with the leading cell uppermost, changes in cell position or number of cells in the chain over time are shown along the x axis. In example A, the chain starts with one fibroblast at the front and one following SCC and grows by the incorporation of two SCC cells at the ‘rear’. The position of cells relative to one another does not change. In example B, the chain grows by the incorporation of three SCC cells and the position of SCC cells relative to each other varies but fibroblasts always remain at the front. In example C, the number of cells in the chain does not change but the position of SCC cells relative to one another varies.
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This note was uploaded on 08/17/2009 for the course 26CB 880 taught by Professor Khan during the Spring '07 term at University of Cincinnati.

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Gaggioli 2007 suppl - S U P P L E M E N TA R Y I N F O R M...

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