Defining Lineage-Specific Membrane Fluidity Signatures that Regulate Adhesion Kinetics

Abstract

SummaryCellular membrane fluidity is a critical modulator of cell adhesion and migration, prompting us to define the systematic landscape of lineage-specific cellular fluidity throughout differentiation. Here, we have unveiled membrane fluidity landscapes in various lineages ranging from human pluripotency to differentiated progeny: (1) membrane rigidification precedes the exit from pluripotency, (2) membrane composition modulates activin signaling transmission, and (3) signatures are relatively germ layer specific presumably due to unique lipid compositions. By modulating variable lineage-specific fluidity, we developed a label-free “adhesion sorting (AdSort)” method with simple cultural manipulation, effectively eliminating pluripotent stem cells and purifying target population as a result of the over 1,150 of screened conditions combining compounds and matrices. These results underscore the important role of tunable membrane fluidity in influencing stem cell maintenance and differentiation that can be translated into lineage-specific cell purification strategy.

Monitoring membrane viscosity in differentiating stem cells using BODIPY-based molecular rotors and FLIM

“…In simple synthetic model membranes biophysical properties are determined by lipid composition 23,24 . There is evidence from mass spectrometry analysis of lipid composition of differentiated cells 2,4 and from generalized polarization studies 1 , that differentiation is accompanied by significant changes of membrane lipid composition and order. We hypothesized that MCSs differentiation should be accompanied by changes in membrane lipids, leading to viscosity changes.…”

“…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 self-renewal 2 .…”

“…There is evidence that the membrane fluidity substantially changes during induced pluripotent stem cells (iPS) differentiation. Generalized polarization monitoring was previously used to detect the rise of membrane rigidity during iPSC differentiation 1 . Furthermore, the results in 1 potentially suggested that membrane rigidification could be transmitted to neighboring cells, resulting in the acceleration of a cells differentiation, in a wave-line fashion.…”

“…Generalized polarization monitoring was previously used to detect the rise of membrane rigidity during iPSC differentiation 1 . Furthermore, the results in 1 potentially suggested that membrane rigidification could be transmitted to neighboring cells, resulting in the acceleration of a cells differentiation, in a wave-line fashion. It was also reported that the viscoelastic properties can predict which subpopulations of undifferentiated mesenchymal stem cells (MSCs) differentiate into osteocytes, and which would turn into adipocytes or chondroblasts.…”

Protein Nanoparticle Osmotic Pressure Modifies Nonselective Permeability of the Blood Brain Barrier by Increasing Membrane Fluidity

“…BBB lesion-factor-induced uidity of occludin decreases after stabilization of MFs or MTs Membrane uidity can have a crucial effect on nonselective permeability of the BBB, which is associated with motion of transmembrane proteins [16,30] . The uorescence recovery after photobleaching (FRAP) test was used to study the changes in mobility ratios of the transmembrane protein occludin.…”