Abstract
Drug discovery efforts have an increasing focus on functional cell-based screening to identify compounds that modulate targets presented in a relevant format. Historically, immortalized cell lines have been used in primary and secondary screens due to their ease of manipulation, transformation, and propagation. However, more researchers are using primary cells that present their drug targets in their natural context. Human primary cell isolation and propagation procedures have become efficient enough to provide these cells in the necessary scale for early stage drug discovery. Adult human stem cells provide an opportunity for investigating multiple pathways of differentiation, development, regeneration, and toxicity using a single cell source and type. Adipose-derived stem cells (ASCs) are an attractive adult human primary stem cell for drug discovery due their abundance in adipose tissue, ease of isolation, and propagation in culture. They can be expanded in high numbers and retain their unique properties to differentiate into multiple lineages. In this chapter, we describe a protocol to identify modulators of human ASC lipogenesis following partial differentiation to adipocytes.
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Gasparri, F., Sola, F., Bandiera, T., Moll, J., and Galvani, A. (2008) High-content analysis of kinase activity in cells. Comb Chem High Throughput Screen 11, 523–36.
Minor, L. K. (2008) Label-free cell-based functional assays. Comb Chem High Throughput Screen 11, 573–80.
Wunder, F., Kalthof, B., Muller, T., and Huser, J. (2008) Functional cell-based assays in microliter volumes for ultra-high throughput screening. Comb Chem High Throughput Screen 11, 495–504.
Goldbard, S. (2006) Bringing primary cells to mainstream drug development and drug testing Curr Opin Drug Discov Devel 9, 110–6.
Dunne, A., Jowett, M., and Rees, S. (2009) Use of primary human cells in high-throughput screens Meth Mol Biol 565, 239–57.
Maddox, C. B., Rasmussen, L., and White, E. L. (2008) Adapting cell-based assays to the high throughput screening platform: problems encountered and lessons learned. JALA Charlottesv Va 13, 168–73.
Crook, J. M. and Kobayashi, N. R. (2008) Human stem cells for modeling neurological disorders: accelerating the drug discovery pipeline. J Cell Biochem 105, 1361–6.
Eglen, R. M., Gilchrist, A., and Reisine, T. (2008) An overview of drug screening using primary and embryonic stem cells. Comb Chem High Throughput Screen 11, 566–72.
Fang, Y. Q., Wong, W. Q., Yap, Y. W., and Orner, B. P. (2007) Stem cells and combinatorial science. Comb Chem High Throughput Screen 10, 635–51.
Nirmalanandhan, V. S. and Sittampalam, G. S. (2009) Stem cells in drug discovery, tissue engineering, and regenerative medicine: emerging opportunities and challenges. J Biomol Screen 14, 755–68.
Zhang, X. Z., Li, X. J., Ji, H. F., and Zhang, H. Y. (2009) Impact of drug discovery on stem cell biology. Biochem Biophys Res Commun 383, 275–9.
Sun, N., Panetta, N. J., Gupta, D. M., Wilson, K. D., Lee, A., Jia, F., Hu, S., Cherry, A. M., Robbins, R. C., Longaker, M. T., and Wu, J. C. (2009) Feeder-free derivation of induced pluripotent stem cells from adult human adipose stem cells. Proc Natl Acad Sci U S A 106, 15720–15725.
Halvorsen, Y. D., Bond, A., Sen, A., Franklin, D. M., Lea-Currie, Y. R., Sujkowski, D., Ellis, P. N., Wilkison, W. O., and Gimble, J. M. (2001) Thiazolidinediones and glucocorticoids synergistically induce differentiation of human adipose tissue stromal cells: biochemical, cellular, and molecular analysis. Metabolism 50, 407–13.
Aust, L., Devlin, B., Foster, S. J., Halvorsen, Y. D., Hicok, K., du Laney, T., Sen, A., Willingmyre, G. D., and Gimble, J. M. (2004) Yield of human adipose-derived adult stem cells from liposuction aspirates. Cytotherapy 6, 7–14.
Zhang, J. H., Chung, T. D., and Oldenburg, K. R. (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 4, 67–73.
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Lea-Currie, Y.R., Duffin, D.J., Buehrer, B.M. (2011). Use of Adipose-Derived Stem Cells in High-Throughput Screening to Identify Modulators of Lipogenesis. In: Gimble, J., Bunnell, B. (eds) Adipose-Derived Stem Cells. Methods in Molecular Biology, vol 702. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61737-960-4_26
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DOI: https://doi.org/10.1007/978-1-61737-960-4_26
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Publisher Name: Humana Press, Totowa, NJ
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