ASGSB 2003 Annual Meeting Abstracts


[58]

GENE ARRAY ANALYSIS OF 3-D CELL AGGREGATES OF MONOCULTURES AND COCULTURES OF TUMOR-DERIVED CELLS.  V. Chopra*, E. V.Hannigan*, B.A. Luxon*, T.G. Wood*, N.R. Pellis**, *UTMB, Galveston, TX and **NASA-JSC, Houston, TX.

     Co-cultures of three-dimensional (3-D) constructs of one cell type with dispersed cells of a second cell type in low-shear rotating suspension cultures in analogue microgravity environment were used to investigate invasive properties of normal and malignant cells that govern tumor angiogenesis.  Various staining procedures and gene array analysis of tumor-derived epithelial cells (TEC) cultured as monocultures in the STLV bioreactor as 3-D cell aggregates; endothelial cells (HUVEC) cultured as monocultures; and also cocultures of TEC with HUVEC angiogenic response of tumors by using probe sets of the human clone HG-U95B analyzed by using Affymatrix gene array analysis.  626 probe sets were differentially expressed by monocultures of HUVEC and 1300 probe sets by monocultures of TEC.  There was a differential expression of genes governing the production of interleukin-2, -6, and 8, vascular endothelial cells in comparison with the cocultures.  There was also a change in expression patterns of 177 genes in the two-dimensional (2-D) regular flask cultures in comparison with the cocultures.  There was also a change in comparison with the 3-D monocultures and cocultures.  The production of these factors was also confirmed by using their respective assays like ELISA and RT-PCR and cytochemical and immunohistochemical staining procedures.  We have shown that the epithelial and endothelial cells undergo a switch in characteristics when grown in an in vitro 3-D environment as compared to their respective monocultures.  This coculture provides new insights into the invasive process and its effects on both invading and invaded cells and can be used to study the eeffectiveness of various antiangiogenic agents.   Funded by NASA.

  

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