Louis T. McLane

Characterization of a multilayer heparin coating for biomolecule presentation to human mesenchymal stem cell spheroids

Mesenchymal stem cells therapies have the potential to treat many pathologies, however, controlling cell fate after implantation remains challenging. We have used a multilayer technology to graft a range of 5 μg/mL - 5 mg/mL heparin onto the surface of MSC aggregates. Heparin coating does not affect cell viability (seen through LIVE/DEAD staining), cell anti-inflammatory properties (seen through co-culture with activated monocytes)and facilitates sequestration by coated cells of a growth factor (TGF-β1) that remains bioactive. This system can maximize therapeutic potential of MSC-based treatments because the cell surface-loaded protein could both signal to the cells to influence transplanted cell fate and be released into the surrounding environment to help repair injured tissue.

Cell Surface Access Is Modulated by Tethered Bottlebrush Proteoglycans

The hyaluronan-rich pericellular matrix (PCM) plays physical and chemical roles in biological processes ranging from brain plasticity, to adhesion-dependent phenomena such as cell migration, to the onset of cancer. This study investigates how the spatial distribution of the large negatively charged bottlebrush proteoglycan, aggrecan, impacts PCM morphology and cell surface access. The highly localized pericellular milieu limits transport of nanoparticles in a size-dependent fashion and sequesters positively charged molecules on the highly sulfated side chains of aggrecan. Both rat chondrocyte and human mesenchymal stem cell PCMs possess many unused binding sites for aggrecan, showing a 2.5x increase in PCM thickness from ∼7 to ∼18 μm when provided exogenous aggrecan. Yet, full extension of the PCM occurs well below aggrecan saturation. Hence, cells equipped with hyaluronan-rich PCM can in principle manipulate surface accessibility or sequestration of molecules by tuning the bottlebrush proteoglycan content to alter PCM porosity and the number of electrostatic binding sites.

Force measurements with a translating holographic optical trap

Force measurements made with a translating holographic optical trap (HOT) of a viscous and a viscoelastic medium are investigated. In purely viscous media, Stokes drag cannot be measured with a translating HOT with established methods. In the viscoelastic system of the pericellular coat, the standard force curves generated by a fixed optical trap coupled with a moving stage can reliably be reproduced by translating HOT experiments. The viscoelastic cell coat provides an example where slow relaxation dynamics makes force measurements relatively insensitive to differences between measurements. These preliminary studies suggest that when the relaxation time scale of a system is much slower than the time scale of the HOT updates, translating HOTs can be reliably used to make force measurements on a viscoelastic, non-equilibrium system.