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Single-cell, high-throughput analysis of cell docking to vessel wall.

J Cereb Blood Flow Metab. 2019 Nov;39(11):2308-2320. doi: 10.1177/0271678X18805238. Epub 2018 Oct 26.

Andrzejewska A1, Nowakowski A1, Grygorowicz T2, Dabrowska S1, Orzel J3,4, Walczak P5,6,7, Lukomska B1, Janowski M1,5,6.

Author information

  1. NeuroRepair Department, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
  2. Laboratory of Advanced Microscopy Techniques, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
  3. Department of Experimental Pharmacology and Laboratory of Nuclear Magnetic Resonance Imaging, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
  4. Faculty of Electronics, Warsaw University of Technology, Warsaw, Poland.
  5. Russell H. Morgan Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
  6. Cellular Imaging Section and Vascular Biology Program, Institute for Cell Engineering, Johns Hopkins University, Baltimore, MD, USA.
  7. Department of Neurology and Neurosurgery, Faculty of Medical Sciences, University of Warmia and Mazury, Olsztyn, Poland.

Abstract

Therapeutic potential of mesenchymal stem cells (MSCs) has been reported consistently in animal models of stroke, with mechanism mainly through immunomodulation and paracrine activity. Intravenous injection has been a prevailing route for MSCs administration, but cell quantities needed when scaling-up from mouse to human are extremely high putting into question feasibility of that approach. Intra-arterial delivery directly routes the cells to the brain thus lowering the required dose. Cell engineering may additionally improve cell homing, further potentiating the value of intra-arterial route. Therefore, our goal was to create microfluidic platform for screening and fast selection of molecules that enhance the docking of stem cells to vessel wall. We hypothesized that our software will be capable of detecting distinct docking properties of naïve and ITGA4-engineered MSCs. Indeed, the cell flow tracker analysis revealed positive effect of cell engineering on docking frequency of MSCs (42% vs. 9%, engineered vs. control cells, p < 0.001). These observations were then confirmed in an animal model of focal brain injury where cell engineering resulted in improved homing to the brain. To conclude, we developed a platform to study the docking of cells to the vessel wall which is highly relevant for intraarterial cell targeting or studies on neuroinflammation.

KEYWORDS:

ITGA4; Mesenchymal stem cells; docking; mRNA; microfluidic assay; stroke

Additional information