Abstract
BackgroundIdentifying and isolating cells with specific behavioral characteristics will facilitate the understanding of the molecular basis regulating these behaviors. Although many approaches exist to characterize cell motility, retrieving cells of specific motility following analysis remains challenging.ResultsCells migrating on substrates coated with fluorescent microspheres generate non-fluorescent tracks as they move and ingest the spheres. The area cleared by each cell allows for quantitation of single cell and population motility; because individual cell fluorescence is proportional to motility, cells can be sorted according to their degree of movement. Using this approach, we sorted a glioblastoma cell line into high motility and low motility populations and found stable differences in motility following sorting.ConclusionWe describe an approach to identify, sort, and enrich populations of cells possessing specific levels of motility. Unlike existing assays of cell motility, this approach enables recovery of characterized cell populations, and can enable screens to identify factors that might regulate motility differences even within clonal population of cells.
Highlights
Identifying and isolating cells with specific behavioral characteristics will facilitate the understanding of the molecular basis regulating these behaviors
Recent advances in tissue microdissection enables highly specific isolation of cells from tissue samples [2], the ability to identify and isolate living cells based on specific behavioral characteristics may provide valuable insights that may not be evident from static morphological analysis of tissue [3,4]
Migrating cells create non-fluorescent tracks on fluorescent microsphere-coated substrates Non-cytotoxic fluorescent polystyrene microspheres have been utilized as cell labels [9], microinjectable cell tracers [10], retrograde neuronal markers [11], and phagocytosis indicators [12]
Summary
Identifying and isolating cells with specific behavioral characteristics will facilitate the understanding of the molecular basis regulating these behaviors. Many developing tissues are comprised of morphologically indistinguishable cells. These cells are often heterogeneous with respect to gene and protein expression, as well as developmental potential. Methods that facilitate the identification and isolation of cells exhibiting specific behaviors may lead to greater understanding of molecular mechanisms underlying cancer progression. Recent advances in tissue microdissection enables highly specific isolation of cells from tissue samples [2], the ability to identify and isolate living cells based on specific behavioral characteristics may provide valuable insights that may not be evident from static morphological analysis of tissue [3,4]
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