Stem Cell Research
Sample Data
Over the last decade, interest has rapidly grown beyond hematopoietic stem cells to understand the role of embryonic and other adult (or somatic) stem cell populations. BD flow cytometry analyzers can be used in combination with fluorochrome-conjugated antibodies to count members of cell subpopulations and obtain data on the relative expression level of multiple markers for individual self-renewing or differentiated cells.
The pluripotency status of human ES (H9) cells analyzed using the BD™ Human Pluripotent Stem Cell Transcription Factor Analysis Kit
Human ES cells were gated after cytometer setup using BD™ CompBead Plus (Figure 1A). Undifferentiated Nanog+Oct3/4+, Oct3/4+Sox2+, and Nanog+Sox2+ human ES cells composed 99%, 99%, and 99% of the cell population, respectively (Figure 1 B,C,D). Histograms displaying cell counts versus the relative expression level of individual markers are also shown (Figure 1 E,F,G).
Differentiation of mESCs to mesendoderm monitored by flow cytometry
Mouse ES cells (ESE14TG2a) were treated with 10 μM retinoic acid for 5 days. Cells were harvested on day 0, day 2, and day 5. Cells were analyzed for pluripotency and differentiation status using the BD Mouse Pluripotent Stem Cell Transcription Factor Analysis Kit. In addition to the pluripotency markers Nanog, Oct3/4, and Sox2, the transcription factor GATA4 was simultaneously analyzed using a GATA4 Alexa Fluor® 488 (Cat. No. 560330) drop-in antibody. After a 5-day treatment with retinoic acid, the percentage of cells expressing Nanog, Oct3/4, and Sox2 markedly decreases while the percentage of cells expressing GATA4 increases.
