Abstract
Membrane fluidity plays an important role in many cell functions such as cell adhesion, and migration. In stem cell lines membrane fluidity may play a role in differentiation. Here we report the use of viscosity-sensitive fluorophores based on a BODIPY core, termed “molecular rotors”, in combination with Fluorescence Lifetime Imaging Microscopy, for monitoring of plasma membrane viscosity changes in mesenchymal stem cells (MSCs) during osteogenic and chondrogenic differentiation. In order to correlate the viscosity values with membrane lipid composition, the detailed analysis of the corresponding membrane lipid composition of differentiated cells was performed by time-of-flight secondary ion mass spectrometry. Our results directly demonstrate for the first time that differentiation of MSCs results in distinct membrane viscosities, that reflect the change in lipidome of the cells following differentiation.
Highlights
Membrane fluidity is considered a key parameter influencing biological function of cells, such as cell adhesion, migration and differentiation[1]
We report the use of fluorescence lifetime imaging microscopy (FLIM) and viscosity-sensitive fluorophores termed “molecular rotors” as an attractive approach that allows the direct measurement of the plasma membrane viscosity in cell culture in a non-destructive manner, while analyzing a large number of cells at the same time[10,11,12,13,14]
We investigated the membrane viscosity changes in stem cells during differentiation on the basis of fluorescence lifetimes of a new viscosity sensitive molecular rotor BODIPY 1, with good aqueous solubility
Summary
Membrane fluidity is considered a key parameter influencing biological function of cells, such as cell adhesion, migration and differentiation[1] These properties are of particular importance in stem cell lines, where small modifications in membrane parameters have the potential to either promote a lineage commitment or a selfrenewal[2]. Little is known on how the viscosity parameters of different stem cell lineages can change depending on the direction of differentiation. We report the use of fluorescence lifetime imaging microscopy (FLIM) and viscosity-sensitive fluorophores termed “molecular rotors” as an attractive approach that allows the direct measurement of the plasma membrane viscosity in cell culture in a non-destructive manner, while analyzing a large number of cells at the same time[10,11,12,13,14]. We have previously successfully applied FLIM with molecular rotors to measure the plasma membrane viscosity, using the rotors that can be targeted to the plasma m embrane[10,11,12]
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