All things cell culture on the μSiM
Introduction Studies have shown that the shear-stimulation of endothelial cells results in the upregulation of the shear-sensitive transcription factor Kruppel-like factor 2 (KLF2). KLF2 plays a critical role in the regulation of endothelial activity such as the regulation of vascular tone, anti-inflammatory responses, and antithrombotic functions. KLF2 is also responsible for the upregulation of endothelial…
Background Tissue-on-a-chip systems have been a promising means of mimicking and understanding the complex and convoluted functions of the in vivo blood-brain barrier (BBB). These semipermeable borders of endothelial cells are highly selective in order to protect the fragile central nervous system (CNS) from bloodborne pathogens or inflammatory signals and are known to have a…
Introduction The blood-brain barrier (BBB) plays an important role in maintaining brain chemistry by restricting the permeability of small molecules and cells. Inflammatory diseases may disrupt the BBB and promote the development of neurological disorders [1]. One potential mechanism of BBB disruption is the degradation of the extracellular matrix proteins that make up the vascular…
Introduction The µSiM BBB model is being developed at UR with the goal to understand the impact of sepsis on the brain. Here, we use an EECM-BMEC-like cell (iSPC-derived brain endothelial cells) [1] monoculture model and an EECM-BMEC-like cell and brain pericyte-like cell (BPLC; iSPC-derived brain pericytes) [2] coculture model to optimize the concentration of…
Introduction High throughput imaging techniques in conjunction with in vitro micro-physiological systems (MPS) allow for comprehensive exploration of physiologically relevant phenomena in a living 3D mimetic of a tissue microenvironment. Prior work in this lab has readily demonstrated that vascular tissue mimetics (the µSiM) facilitate real time phase-microscopy visualization of immune cell trafficking in response…
Introduction The blood-brain barrier (BBB) plays an important role in maintaining brain chemistry by restricting the permeability of small molecules and cells. Inflammatory diseases may disrupt the BBB and promote the development of neurological disorders [1]. One potential mechanism of BBB disruption is the degradation of the extracellular matrix proteins that make up the endothelial…
One of the validations of the in vitro human Blood Brain Barrier system is to ensure that the endothelial cells derived from iPSC’s and cultured in our system exhibit behaviors that are consistent with endothelial cell behavior in situ. The particular aspect of BBB endothelial function addressed in this post is the expression and properties…
Introduction In the Glading Lab we study Cerebral Cavernous Malformations (CCMs). In CCMs, brain capillaries develop enlarged lumens and a disrupted blood-brain barrier. The presence of a mutation in KRIT1 causes loss of KRIT1 protein function in the endothelial cells. KRIT 1 plays a role in the integrity of endothelial junctions, therefore the lack of…
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…