Introduction Pericytes are mural cells that line the outer surface of microvessels and regulate diverse aspects of vascular development and function. We differentiate brain-pericyte-like cells (BPLCs) from hiPSC clones using the protocol published by Eric Shusta’s lab (Gastfriend et al., 2021) (Figure 1). This protocol describes methods to differentiate hiPSCs to BPLCs via neural crest…
Introduction We are designing a cell culture chamber insert to allow side-by-side co-culture of cells and ultimately triculture in the µSiM devices. Materials and Methods ALine devices assembly according to the “ALine Modular Device Assembly” instructions. 2-slot NPN membranes applied (NPSN100-2L, nanoporous silicon nitride 100 nm thick, 2 windows). Straight tweezers and chip tweezers are…
In previous experiments where the affinity based capture of EVs in ALine devices was tested, issues were encountered. It was noticed that when capturing EVs on the ALine, the non-biotinylated slit membranes had captured a higher quantity of EVs compared to biotinylated membranes. This conclusion led us to question if the flow through the ALine…
Introduction Risks of extracorporeal membrane oxygenation (ECMO) therapy can be attributed to large membrane areas and blood circuit volumes. These are particularly problematic in neonates with low blood volume. To address this, we are developing a miniaturized prototype microfluidic ECMO using ultrathin highly porous silicone membranes. Hemocompatibility (hemolysis and thrombosis) assessment of geometrical variants of…
Introduction We are investigating some issues seen with long term attachment (up to 6 days) of the EECM-BMEC-like cells seen by collaborators in Bern, where they noticed black aggregates on the coated control devices that get worse at day 6. Some attachment issues and aggregates have also been seen at the University of Rochester in…
Introduction The work in this post explores the hypothesis that durotaxis affects the invasion of S. aureus into the osteocyte lacuna-canicular network (OLCN). To address this hypothesis, we modified the µSiM to enable the independent tuning of pore stiffness, ranging from those of soft-tissues to that of silicon nitride (~10X stiffer than bone). More background…
“I’ve looked at clouds from both sides now” is a Joni Mitchell song. Some Background We are developing a white blood cell (WBC) detector to surveil peritoneal dialysate fluid during fluid exchanges for early detection of peritonitis. This preemptive monitoring is currently not available in clinical practice. The current detection method is by visual inspection…
As discussed in a previous post of mine, I am currently working on a project to develop an in vitro mimetic of the osteocyte lacuna-canicular network (OLCN) to improve the ability to predict Staphylococcus Aureus invasion and colonization of the OLCN during osteomyelitis. The purpose of this post is to provide more background behind Aim…
Introduction The invasion of Staphlyococcus aureus into the cracks of cortical bone leads to osteomyelitis, a devastating disease often resolved by amputation [1]. The μSiM platform has been used to study the genetic mechanisms of S. aureus invasion. The membrane pores mimic the network of cracks in bone, referred to as the osteocyte lacuno-canalicular network…
Introduction Extracorporeal membrane oxygenation (ECMO) devices are used to supply oxygen to patients when they are unable to get sufficient oxygen through their lungs. Miniaturizing an ECMO device could be beneficial for neonatal patients, as it would require less blood to prime the device and operate efficiently at a lower overall flow volume. Dialysis is…