Dmytriyev Anatoliy, Tkach Vadim, Rudenko Olga, Bock Elisabeth, Berezin Vladimir
Protein Laboratory, Institute of Molecular Pathology, Copenhagen University, Copenhagen, Denmark.
Cytometry A. 2006 Sep 1;69(9):979-85. doi: 10.1002/cyto.a.20303.
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.
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.
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.
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).
细胞运动在许多生理和病理过程中至关重要，如胚胎发生、炎症、伤口愈合和转移。然而，评估单个细胞运动时耗时的步骤通常是分析数百张细胞培养的记录图像，包括一次检索一张图像，并用计算机鼠标标记单个细胞的位置。因此，本研究的目的是开发一种用于评估细胞运动的新型自动程序。
该程序在通过瞬时转染或腺病毒转导工程化表达绿色荧光蛋白的成纤维细胞、胶质瘤细胞和腺癌细胞上进行测试，从而能够自动识别检索图像上的细胞坐标。
确定了血清生长因子、致畸化合物和转录因子过表达对培养细胞运动行为的影响。通过手动和自动标记细胞位置来估计细胞运动，随后计算运动参数。发现两种程序获得的结果具有显著相关性。
我们开发了一种程序，可对用表达增强型绿色荧光蛋白（EGFP）的质粒转染或用重组病毒转导的稀疏接种细胞进行自动视频记录。
