Multispectral characterisation of Mesenchymal Stem/ Stromal Cells: Age, Cell cycle, Senescence, and Pluripotency

摘要

Multispectral assessment of cell autofluorescence gives a direct window into the molecular processes occurring withinthose cells. This can be used to non-invasively characterise and classify various cellular properties without requiringfixation, dyes or transformation. Human mesenchymal stem/ stromal cells (MSCs) have great potential to contributeto regenerative medicine, especially with regards to autologous transplantation. However, this capacity is oftenlimited by inherent properties of cell lines, which prevent their being sufficiently expanded after derivation foreffective clinical application. The investigation of these properties requires numerous, time and labour-intensiveassays. In this study we have used correlative microscopy based on multispectral images of cell autofluorescencethen correlated to functional assays in order to construct multispectral signatures of numerous inherent cellcharacteristics. These included cell cycle status (indicating the proportion of cells undergoing cell division at agiven time), cell ‘age’ (number of passages undergone, indicating capacity for further expansion), and β-galactosidase (a marker of senescence, indicating cells which can no longer divide). This study has established asingle protocol, in place of multi-functional assays, to characterize the growth and differentiation capacity of hMSClines using a non-invasive approach.