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Supplemental Digital Content 1. Isotype. CD44. CD45. CD14. CD11b. A. BMMC. B. Nonadherent Cells Passage 0. C. Adherent MSC Passage 0. D. Adherent MSC Passage 1. E. Adherent MSC Passage 2.
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Supplemental Digital Content 1 Isotype CD44 CD45 CD14 CD11b A BMMC B Nonadherent Cells Passage 0 C Adherent MSC Passage 0 D Adherent MSC Passage 1 E Adherent MSC Passage 2 Figure 1. Analysis of canine BMMC and MSC cultures at passages 0 to 2 for expression of cell surface antigens using canine-specific antibodies against CD44, CD45, CD14 and CD11b. Flow cytometric analysis of cell surface markers expressed by (A) noncultured BMMC, (B) the nonadherent fraction from MSC cultures at passage 0, and adherent MSC at (C) passage 0, (D) passage 1, and (E) passage 2. Antibodies used were (from left to right): isotype control, cCD44, cCD45-biotin, cCD14, cCD11b, with FITC-labeled secondary antibodies (mIgG2a for CD44, and mIgG1 for CD14 and CD11b) and Streptavidin-FITC for CD45-biotin. Secondary-only controls were used to set the limits for positive gates. With passage, the proportion of hematopoietic cells within the adherent cell fractions decreased to <1% while hematopoietic cells were found within the nonadherent cell fraction.