Characterization of the KG1a cell line for use in a cell migration based screening assay |
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Authors: | Karl Francis Gyun Min Lee Bernhard O. Palsson |
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Affiliation: | (1) Department of Bioengineering, University of California San Diego, 9500 Gilman Drive Mail Code 0412, 92093-0412 La Jolla, CA, USA;(2) Department of Biological Sciences, KAIST, 373-1 Kusong-dong, Yusong-gu, 305-701 Daejeon, Korea |
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Abstract: | High-throughput screening has become a popular method used to identify new “leads” for potentially therapeutic compounds. Further screening of these lead compounds is typically done with secondary assays which may utilize living, functioning cells as screening tools. A problem (or benefit) with these cell-based assays is that living cells are very sensitive to their environment. We have been interested in the process of stem cell migration and how it relates to the cellular therapy of bone marrow transplantation. In this study we describe a secondary, cell-based assay for screening the effects of variousin-vitro conditions on Immature Hematopoietic Cell (IHC) migration. Our results have revealed many subtle factors, such as the cell's adhesive characteristics, or the effect of a culture's growth phase, that need to be accounted for in a screening protocol. Finally, we show that exponentially growing KG1a cells (a human IHC cell line) were 10 times more motile than those in the lag or stationary phases. These data strongly suggest that KG1a cells secrete a chemokinetic factor during the exponential growth phase of a culture. |
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Keywords: | cell motility hematopoietic stem cells time lapse microscopy cell migration |
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