Journal: EBioMedicine

Article Title: Modulating endothelial adhesion and migration impacts stem cell therapies efficacy

doi: 10.1016/j.ebiom.2020.102987

Figure Lengend Snippet: Human MSC migration in vitro and their adhesion and migration in the lung in vivo (a) Migration capacity expressed by % of untreated MSC and PEI treated MSC with or without additional chemoattractants or human platelet lysate (HPL). Migration data of untreated and PEI treated MSC were compared for each condition with two-tailed t -test (* p < 0.05), N = 3, each data point represents one biological replicate per group Error bars: SD. (b) Immunofluorescence and associated quantification of MSC accumulation in the lung after intravenous application of 5 × 10 6 treated or untreated eGFP+ MSC in a rat model of unilateral hippocampal quinolinic acid (QA)-induced brain injury. Animals were sacrificed 24 h after application. Untreated and treated MSC were detected in the lumina of the small vessels (arrows), but also in close vicinity to the vessels (arrowheads). von Willebrand factor (vWF): red; GFP: green; DAPI: blue; scale bars: 100 μm. Intraparenchymal and intravascular cell numbers were compared for untreated MSC, PEI treated MSC or PEI-NP treated MSC with two-tailed unpaired t -test (*** p < 0.001). N = 3 per group. Intravascular data represent 9 data points generated from 3 sections per mouse ( n = 3). Intraparenchymal data represent 80 –180 data points generated from 6 to 9 micrographs from 8 to 30 sections. Error bars: SEM. (c) Numbers of untreated MSC, PEI-MSC or PEI-NP-MSC detected in the vasculature and parenchyma. Cell numbers of untreated or PEI/NP treated MSC were compared for intraparenchymal or intravascular localisation with ANOVA followed by Tukey's multiple comparisons test (* p < 0.05, *** p < 0.001). N = 3 per group. Intravascular data represent 9 data points generated from 3 slides from N = 3 mice; intraparenchymal data represent 180 data points obtained from 30 slides per mouse (6 micrographs per section). Error bars: SEM. (d) Immunofluorescence of CCL2 and SDF-1 in the pulmonary vasculature and parenchyma of rats administered with untreated MSC, PEI-MSC or PEI-NP-MSC. CCL2: red; GFP: green; DAPI: blue; scale bars: 100 μm SDF-1: red; GFP: green; DAPI: blue; scale bars: 50 μm (MSC, PEI-MSC), 100 μm (PEI-NP-MSC). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Brain and lung sections of QA-lesioned rats, selected from the vicinity of the sections containing human eGFP+BM-MSC, were fixed with methanol at −20 °C, washed and subsequently incubated with antibodies against von Willebrand factor (vWF) diluted 1:300 (rabbit polyclonal, DakoCytomation, Glostrup, Denmark), or SDF-1 diluted 1:50 (rabbit polyclonal, Abcam), or CCL2 diluted 1:200 (rabbit polyclonal, Biozol, Eching, Germany) for 2 h at room temperature (RT).

Techniques: Migration, In Vitro, In Vivo, Two Tailed Test, Immunofluorescence, Generated