An automatic procedure for evaluation of single cell motility.
BACKGROUND
Cell motility is vital in many physiological and pathological processes, such as embryogenesis, inflammation, wound healing, and metastasis. However, the time-consuming step in the evaluation of individual cell motility is the analysis of hundreds of recorded images of cell cultures in general consisting of retrieving images, one at a time, and marking the positions of individual cells by a computer mouse. Therefore, the aim of the present study was to develop a novel automatic procedure for the evaluation of cell motility.


METHODS
The procedure was tested on fibroblasts and glioma and adenocarcinoma cells engineered to express the green fluorescent protein by either transient transfection or adenovirus transduction, allowing automatic recognition of cell coordinates on retrieved images.


RESULTS
The effects of serum growth factors, teratogenic compounds, and overexpression of transcription factors on the motile behavior of cultured cells were determined. Cell motility was estimated by both manual and automatic marking of cell position and subsequently motility parameters were computed. The results obtained by the two procedures were found to correlate significantly.


CONCLUSIONS
We developed a procedure allowing automatic video recording of sparsely seeded cells transfected with a plasmid or tranduced with a recombinant virus expressing the enhanced green fluorescent protein (EGFP).
Dmytriyev
Anatoliy
A
Protein Laboratory, Institute of Molecular Pathology, Copenhagen University, Copenhagen, Denmark.
Tkach
Vadim
V
Rudenko
Olga
O
Bock
Elisabeth
E
Berezin
Vladimir
V
D000230:Adenocarcinoma; D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005910:Glioma; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008433:Mathematics; D051379:Mice; D018715:Microscopy, Video / Q000379:methods*; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D051381:Rats; D013723:Teratogens / Q000494:pharmacology; D014635:Valproic Acid / Q000494:pharmacology; D000099169:Fos-Related Antigen 1
D004791:Enzyme Inhibitors; D049452:Green Fluorescent Proteins; D016760:Proto-Oncogene Proteins c-fos; D013723:Teratogens; D014635:Valproic Acid; D000099169:Fos-Related Antigen 1