Analysis of Breast Cell-Lineage Response Differences to Taxol Using a Novel Co-Culture System
Final rept. 1 Apr 2002-31 May 2005
TEXAS TECH UNIV LUBBOCK
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We have established a new co-culture system in which human mammary epithelial cells HMEC and human mammary tumor cells HMT are physically grown together. We hypothesized that cells in co-culture CC would generate gene expression profiles different from homogeneous cell populations. In this study, cells were incubated with blue or red CellTracker Dyes and co-cultured. Our novel capture system allowed cells to be co-cultured and then quickly separated while maintaining 90 viability. Using deconvolution microscopy, co-cultured HMEC were observed to form focal, gland-like structures surrounded by TTUderived from an invasive ductal carcinoma. Using a differential trypsinization technique, cell populations were rapidly separated to perform RNA extractions performed 2 h in order to obtain expression profiles from still viable cells. RT2 profiler PCR Array SuperArray RT-PCR was utilized to analyze differential gene expression between parent cell lines and cells co-cultured. We observed that in co-culture HMEC become more cancerous compared to homogenous parent HMEC and CC - TTUexhibit gene expression profiles considered to be less cancerous than that of parent HMT. Replated CC HMEC have both gene expression profiles of CC HMEC and parent HMEC, but were more similar to CC HMEC. Replated TTU showed similar results.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research