Abstract
Brain-infiltrating monocyte-derived macrophages are one of the key players in the local immune response after stroke. It is now widely accepted that the inflammatory response is not an exclusively destructive process. However, the underlying molecular mechanisms needed for proper regulation still remain to be elucidated. Here, we propose an in vitro labelling strategy for multimodal in vivo observation of macrophage dynamics distinguished from brain-residing microglia response. Prior to intracerebral transplantation into the striatum of recipient mice or systemic administration, monocytes and macrophages, isolated from luciferase-expressing mice, were labelled with superparamagnetic iron oxide particles. Temporo-spatial localization was monitored by magnetic resonance imaging, whereas survival of grafted cells was investigated using bioluminescence imaging. The labelling procedure of the isolated cells did not significantly influence cell characteristics and resulted in detection of as few as 500 labelled cells in vivo. Two weeks after stereotactic transplantation, the luciferase signal was sustained traceable, with approximately 18% of the original luciferase signal detectable for monocytes and about 30% for macrophages. Hypointensity in MRI of the graft appeared unaltered in spatial location. In a therapeutically relevant approach, systemic cell administration after stroke resulted in accumulation mostly in thoracic regions, as could be visualized with BLI. For detection of homing to ischemic brain tissue more cells need to be administered. Nevertheless, during parallel MRI sessions recruitment of i.v. injected cells to the lesion site could be detected by day 2 post stroke as scattered hypointense signal voids. With further increase in sensitivity, our multi-facetted labelling strategy will provide the basis for in vivo tracking and fate specification of tissue-infiltrating macrophages and their distinct role in stroke-related neuro-inflammation.
Highlights
Ischemic or traumatic brain injuries or other cerebral diseases are accompanied by a strong local inflammatory response in the affected tissue [1, 2]
Well-established protocols combined with the use of contrast agents allow for investigation of the spatial localization of implanted cells in the host brain tissue [13, 14]
We show that the efficient labelling strategy based on concatenation of the use of luciferase-expressing transgenic cells and physical labelling with iron oxide nanoparticles was successfully established in vitro and validated for the influence on cell functionality
Summary
Ischemic or traumatic brain injuries or other cerebral diseases are accompanied by a strong local inflammatory response in the affected tissue [1, 2]. It has been suggested that the monocytes play a regulatory role in the inflammation process [4,5,6,7, 11, 12] with the essential function of co-localization with the glial scar. This step is crucial for proper scar resolution and termination of the local microglial response. Well-established protocols combined with the use of contrast agents (superparamagnetic particles of iron oxide–SPIOs) allow for investigation of the spatial localization of implanted cells in the host brain tissue [13, 14]. In the present study we I) carefully assessed optimal labelling strategies with SPIO particles for minimal or no label influence on monocytes and MF in vitro; II) investigated labelling effectiveness and detection sensitivity in vitro and in vivo; III) determined longevity of contrast agent and spatial cell localization in a qualitative manner (SPIO-MRI); IV) monitored spatial cell localization longitudinally (SPIO-MRI) and assessed graft survival via bioluminescence imaging (BLI); and, V) applied our findings to experimental stroke, in order to image the recruitment time profile and fate of systemically administered monocytes and MF in vivo (SPIO-MRI and BLI)
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