Primer insan beyni mural hücrelerinin izolasyonu ve karakterizasyonu

Pericytes and smooth muscle cells (SMCs) are mural cells that are located on the abluminal side of the blood vessels. In the central nervous system (CNS), pericytes are implicated to play roles in maintaining the integrity of the blood brain barrier (BBB), and regulation of cerebral blood flow. Recent studies show that pericytes also have immunoregulatory functions. In response to inflammatory stimuli, pericytes have been shown to upregulate expression of cell adhesion molecules such as VCAM-1, ICAM-1 and pro-inflammatory cytokines and chemokines. In genetically engineered animal models, loss of pericytes lead to increased BBB permeability and an increased immune cell infiltration into the CNS. On the other hand, studies investigating the role of pericytes in the tumor microenvironment (TME) showed that tumor associated pericytes could support tumor progression by immunosuppression. Hence, investigating the pathways pericytes use to interact with the immune system can reveal disease mechanisms that could be potential targets for immunomodulatory treatments. However, studying mural cells in vivo and in vitro is difficult as pericytes, SMCs and fibroblasts have a very similar molecular signature. In this study, we aimed to investigate the immunologic properties of brain pericytes by (i) generating primary human brain pericyte cells from tissue obtained during surgery by using two different methods present in the literature, (ii) comparing their molecular characteristics to commercially available human brain pericyte and human brain fibroblast cell lines, (iii) establishing their gene expression pattern changes in response to inflammatory cytokines, (iv) studying human brain pericyte – T lymphocyte interactions in vitro via chemotaxis experiments and (v) generating a CXCL10 knock-out human brain pericyte cell line via CRISPR/Cas9 to study the effect of CXCL10 on human brain pericyte – T lymphocyte chemotaxis. Overall, our data points to a potential pro-inflammatory role for brain pericytes; thus, understanding the specific pathways they are involved in T lymphocyte recruitment into the CNS could help us determine if these pathways can be targeted for interventions in neuroinflammatory conditions.