Quantitative Cell Biology

This project is focused on developing materials and methods that aid the quantitative analysis of cell biology and help minimize sources of variability in the microscopic evaluation of cells and tissues. Based on self-assembly and alkanthiol monolayers, thin films of extracellular matrix proteins provide an analytically validated matrix on which cells can be reproducibly cultured. Thin film materials of extracellular matrix proteins and protocols for quantitative automated fluorescence microscopy and image analysis facilitate measurements of the statistically relevant numbers of cells in an unbiased fashion. Also in development are genetically engineered indicator cells that provide a rapid, nearly real-time readout of cellular response to environment. These tools will facilitate applications such as diagnostic assays, development of biomaterials, determination of culture conditions for stem cells, drug screening and drug development, and validation and use of data from basic research on cells and tissues. Such reference materials and protocols are essential for standardizing and validating data and modeling for understanding complex biological systems.

• To meet industrial needs, CSTL has developed and built expertise in the following areas: cell-biomaterial interactions; reference ECM surfaces; formaldehyde-free fixation for maintaining cellular GFP; quantification of cell morphology and measurement of intracellular Green Fluorescent Protein; quantifying the natural variability within cell populations; validation of indicator cells; live cell imaging; cellular toxicity measurements; and cryopreservation analysis.
• Validated that highly reproducible and homogeneous thin films of the extracellular matrix protein, Type 1 collagen, provide a relevant growth environment to cells, and compare favorably to poorly reproducible alternative collagen matrices.
• Confirmed the biological response of engineered indicator cells containing a gene construct that reliably provides a green fluorescent signal when cells are in a proliferative state.
• Quantitatively analyzed the statistical correlation between multiple intracellular parameters on a cell by cell basis, to understand the relationship between average response of a population of cells and the nature of biological variability from cell to cell.
• The Symposium on Metrology and Standards for Cell Signaling: Impact on Tissue Engineering was held in October 2003 and was co-sponsored by NIST and ASTM International. Renowned experts discussed the following topics: the complexity of cell signaling; examples of directing cell signaling and differentiation; the state of the art in measurement technologies; the collection, treatment, and analysis of cell signaling data; and examples of how knowledge of cell signaling can assist in the design and testing of materials for tissue engineering. http://www.cstl.nist.gov/biotech/cellsignaling/index.html

• A.L. Plant, E. Horowitz, �Symposium on Metrology and Standards for Cell Signaling: Impact on Tissue Engineering� Tissue Eng. , in press.
• J. T. Elliott, A. Tona, A. L. Plant (2002) � Comparison of reagents for shape analysis of fixed cells by automated fluorescence microscopy, � Cytometry 2002, 52A, 90-100 .
• J.T. Elliott, K.J. Langenbach , A. Tona , P.L. Jones, A.L. Plant, �Vascular smooth muscle cell response on thin films of collagen, � Matrix Biol., submitted.
• J. T. Elliott, P. L. Jones, J. T. Woodward, A. Tona , A.L. Plant, �Morphology of smooth muscle cells on thin films of collagen� Langmuir 2002, 19, 1506-1514.
• R. Chapados , D. McKean, L. Sisbarro , J. Elliot, A. Plant, P.L. Jones, �Extracellular matrix-regulated expression of tenascin -C in vascular smooth muscle cells is cell shape-dependent� , Circ. Res., submitted.
• J. T. Elliott, A. Tona , J. T. Woodward, C. W. Meuse , H. M. Elgendy , A.L. Plant, �Osteoblast Cell Membrane Hybrid Bilayers for Studying Cell-Cell Interactions,�  IEEE Procedings: Nanobiotechnology 2004.
• H. Rodriguez, C. O'Connell, P. E. Barker, D. H. Atha, P. Jaruga, M. Birincioglu, M. Marino, P. McAndrew, M. Dizdaroglu, �Measurement of DNA Biomarkers for the Safety of Tissue-Engineered Medical Products Using Artificial Skin as a Mode,� Tissue Eng. 2004, 10, 1332-1345.
• J.Y. Wong, A. L. Plant, editors, �Architecture and Application of Biomaterials and Biomolecular Materials ,� Materials Research Society, 2004.
• K. J. Langenbach, J. T. Elliott, A. Tona, A. L. Plant, �Evaluating the correlation between fibroblast morphology and promoter activity on thin films of extracellular matrix proteins ,� J.Cell Sci., submitted.