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Sci Rep ; 4: 5942, 2014 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-25096726

RESUMO

Quantification of basic cell functions is a preliminary step to understand complex cellular mechanisms, for e.g., to test compatibility of biomaterials, to assess the effectiveness of drugs and siRNAs, and to control cell behavior. However, commonly used quantification methods are label-dependent, and end-point assays. As an alternative, using our lensfree video microscopy platform to perform high-throughput real-time monitoring of cell culture, we introduce specifically devised metrics that are capable of non-invasive quantification of cell functions such as cell-substrate adhesion, cell spreading, cell division, cell division orientation and cell death. Unlike existing methods, our platform and associated metrics embrace entire population of thousands of cells whilst monitoring the fate of every single cell within the population. This results in a high content description of cell functions that typically contains 25,000 - 900,000 measurements per experiment depending on cell density and period of observation. As proof of concept, we monitored cell-substrate adhesion and spreading kinetics of human Mesenchymal Stem Cells (hMSCs) and primary human fibroblasts, we determined the cell division orientation of hMSCs, and we observed the effect of transfection of siCellDeath (siRNA known to induce cell death) on hMSCs and human Osteo Sarcoma (U2OS) Cells.


Assuntos
Fibroblastos/fisiologia , Células-Tronco Mesenquimais/fisiologia , Microscopia de Vídeo/métodos , Osteoblastos/metabolismo , Gravação em Vídeo/métodos , Adesão Celular , Contagem de Células , Morte Celular/genética , Divisão Celular , Linhagem Celular Tumoral , Fibroblastos/citologia , Humanos , Células-Tronco Mesenquimais/citologia , Microscopia de Vídeo/instrumentação , Osteoblastos/patologia , Cultura Primária de Células , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Gravação em Vídeo/instrumentação
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