American Society of Hematology

Figure 3

$Figure 3. SSEA-4 marks murine MSCs. Time course for SSEA-4 and CD45 expression in BM adherent cells by flow cytometry. (A) A representative FACS profile of BM adherent cells analyzed for SSEA-4 and CD45 expression at 2, 4, 7, 14, 21, and 28 days in culture. Fluorescence intensity of SSEA-4 (PE-conjugated) is indicated on the x-axis, and CD45 (APC-conjugated) is shown on the y-axis. Percentages represent the fraction of cells that express a given surface antigen. (B) SSEA-4 and CD45 expression over time at days 2, 4, 8, 15, and 22. The numbers in parentheses below the bars represent the number of independent bone marrow cultures that were assayed at that time point. Error bars indicate standard error. (C) Two days after BM was harvested, adherent BM cells were sorted for SSEA4. Although SSEA4+ and SSEA-4− cells were plated at the same cell density, SSEA-4− cells failed to grow (left), whereas SSEA4+ cells demonstrated extensive growth (right). (D) Expanded SSEA4+ cells were capable of differentiating along fat and bone lineages when induced with appropriate media, as observed by Oil Red O (top left) and Alizarin Red S staining (top right), respectively, as well as by gene expression analyses (shown below). Original magnification for panels C and D, 10×/0.3 NA Ph1.$

SSEA-4 marks murine MSCs. Time course for SSEA-4 and CD45 expression in BM adherent cells by flow cytometry. (A) A representative FACS profile of BM adherent cells analyzed for SSEA-4 and CD45 expression at 2, 4, 7, 14, 21, and 28 days in culture. Fluorescence intensity of SSEA-4 (PE-conjugated) is indicated on the x-axis, and CD45 (APC-conjugated) is shown on the y-axis. Percentages represent the fraction of cells that express a given surface antigen. (B) SSEA-4 and CD45 expression over time at days 2, 4, 8, 15, and 22. The numbers in parentheses below the bars represent the number of independent bone marrow cultures that were assayed at that time point. Error bars indicate standard error. (C) Two days after BM was harvested, adherent BM cells were sorted for SSEA4. Although SSEA4⁺ and SSEA-4⁻ cells were plated at the same cell density, SSEA-4⁻ cells failed to grow (left), whereas SSEA4⁺ cells demonstrated extensive growth (right). (D) Expanded SSEA4⁺ cells were capable of differentiating along fat and bone lineages when induced with appropriate media, as observed by Oil Red O (top left) and Alizarin Red S staining (top right), respectively, as well as by gene expression analyses (shown below). Original magnification for panels C and D, 10×/0.3 NA Ph1.

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