Journal
JOURNAL OF FLUORESCENCE
Volume 24, Issue 5, Pages 1531-1536Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10895-014-1439-3
Keywords
Lung cancer; Orthotopic; Circulating tumor cells; CTC; In vitro culture; CTC; MetaCell; Filtration; Size; Fluorescence; RFP
Funding
- University Hospital Motol, Prague Czech Republic [000 64203]
- Charles University in Prague [P 27/2012]
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In the present study, we demonstrate an animal model and recently introduced size-based exclusion method for circulating tumor cells (CTCs) isolation. The methodology enables subsequent in vitro CTC-culture and characterization. Human lung cancer cell line H460, expressing red fluorescent protein (H460-RFP), was orthotopically implanted in nude mice. CTCs were isolated by a size-based filtration method and successfully cultured in vitro on the separating membrane (MetaCellA (R)), analyzed by means of time-lapse imaging. The cultured CTCs were heterogeneous in size and morphology even though they originated from a single tumor. The outer CTC-membranes were blebbing in general. Abnormal mitosis resulting in three daughter cells was frequently observed. The expression of RFP ensured that the CTCs originated from lung tumor. These readily isolatable, identifiable and cultivable CTCs can be used to characterize individual patient cancers and for screening of more effective treatment.
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