Computational protocol: Intravenous administration of mesenchymal stem cells prevents angiotensin II-induced aortic aneurysm formation in apolipoprotein E-deficient mouse

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Protocol publication

[…] BM-MSCs were isolated from femurs and tibiae of male apoE−/− mice (6 to 8 weeks old) and expanded as described []. The mesenchymal population was isolated on the basis of its ability to adhere to the culture plate. At 90% confluence, the cells were trypsinized (0.05% Trypsin containing 0.53 mM EDTA-4Na, Invitrogen) and were passaged to 175-cm2 flasks. Expansion culture continued up to passages 7–10 were used in all experiments. Cultured BM-MSCs were incubated with the following antibodies, conjugated with phycoerythrin (PE), allophycocyanin (APC) or peridinin chlorophyll protein-Cy5.5 (PerCP-Cy5.5) (eBioscience): CD11b, CD31, CD34, CD44, CD45, CD73, CD106, CD117, and Sca-1. Labeled cells were assayed by flow cytometer (FACS Canto II, Becton Dickinson) and analyzed with BD FACSDiva Software and FlowJo (Tree Star Inc.). The capacity to differentiate into osteogenic, adipogenic, and chondrogenic lineages by addition of inductive media was confirmed with chemical staining as performed previously (Additional file ) []. […]

Pipeline specifications

Software tools BD FACSDiva, FlowJo
Application Flow cytometry
Organisms Mus musculus
Diseases Aortic Aneurysm