The analysis of the movement of the genetically modified human skin fibroblasts in culture

摘要

The aim of this study is the comparison of the movement of various types of human dermal fibroblasts with and without genetic modification. Three groups of fibroblasts were cultured and monitored: (1) control group of fibroblasts isolated from human skin (not transduced); (2) transduced with lentivirus bearing EGFP fluorescent marker; (3) transduced with lentivirus bearing DsRed2 fluorescent marker. The experimental sequences of images documenting cells' movement have been analyzed using image processing methods. The cells' movement was described by: (1) distance between each two consecutive images/frames, (2) distance and displacement covered by each fibroblast in 30 minutes (3) length of cell crawling cycle, (4) the movement tortuosity coefficient for all fibroblasts in each group. Also shape features such as: area, perimeter, eccentricity, length of the major axis have been analyzed. It appears that the general movement behavior is not changed by the process of transduction but some of its aspects are modified. The efficiency of movement in the sense of distance covered and region penetration is decreased because of changes in cells' morphology. Transduced cells are less polarized and develop extra podia during their crawling.