We are working to develop flow-based methods for separating different cell types from a mixed-cell co-culture. We are currently pursuing two separate projects. In the first, we are co-culturing normal fibroblasts with transformed fibroblasts in order to determine if cell-cell interactions can overcome the growth inhibition which the normal cells experience in 3-D culture. In collaboration with Dr. Wright, we have developed subclones of each fibroblast cell type which are stably transfected with green fluorescent protein (GFP). We have characterized the stability of GFP expression in these cells using flow analysis, and are currently conducting experiments using simultaneous flow measurement of GFP and DNA content in order to measure the proliferative status of each cell type in a mixed-cell co-culture. The second project involves investigating the radiation response of monolayer and spheroid co-cultures of normal human prostate and fibroblast cells. We are inv esti gating several potential methods for using flow analysis and sorting to separate these cell types, including fuorescently-tagged antibodies directed against the cell surface, membrane labeling using PKH-dyes, and GFP transfection. New antibodies will be developed using phage display technology. Flow analysis will be critical in evaluating these methods and in isolating high-affinity antibodies for physical separation of the cells from a mixed-cell co-culture by the use of magnetic beads.
| Frumkin, Jesse P; Patra, Biranchi N; Sevold, Anthony et al. (2016) The interplay between chromosome stability and cell cycle control explored through gene-gene interaction and computational simulation. Nucleic Acids Res 44:8073-85 |
| Johnson, Leah M; Gao, Lu; Shields IV, C Wyatt et al. (2013) Elastomeric microparticles for acoustic mediated bioseparations. J Nanobiotechnology 11:22 |
| Micheva-Viteva, Sofiya N; Shou, Yulin; Nowak-Lovato, Kristy L et al. (2013) c-KIT signaling is targeted by pathogenic Yersinia to suppress the host immune response. BMC Microbiol 13:249 |
| Ai, Ye; Sanders, Claire K; Marrone, Babetta L (2013) Separation of Escherichia coli bacteria from peripheral blood mononuclear cells using standing surface acoustic waves. Anal Chem 85:9126-34 |
| Sanders, Claire K; Mourant, Judith R (2013) Advantages of full spectrum flow cytometry. J Biomed Opt 18:037004 |
| Cushing, Kevin W; Piyasena, Menake E; Carroll, Nick J et al. (2013) Elastomeric negative acoustic contrast particles for affinity capture assays. Anal Chem 85:2208-15 |
| Piyasena, Menake E; Austin Suthanthiraraj, Pearlson P; Applegate Jr, Robert W et al. (2012) Multinode acoustic focusing for parallel flow cytometry. Anal Chem 84:1831-9 |
| Austin Suthanthiraraj, Pearlson P; Piyasena, Menake E; Woods, Travis A et al. (2012) One-dimensional acoustic standing waves in rectangular channels for flow cytometry. Methods 57:259-71 |
| Vuyisich, Momchilo; Sanders, Claire K; Graves, Steven W (2012) Binding and cell intoxication studies of anthrax lethal toxin. Mol Biol Rep 39:5897-903 |
| Chaudhary, Anu; Ganguly, Kumkum; Cabantous, Stephanie et al. (2012) The Brucella TIR-like protein TcpB interacts with the death domain of MyD88. Biochem Biophys Res Commun 417:299-304 |
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