Stem Cell Research
Sample Data
The ability to sort pluripotent stem cells, such as human embryonic stem cells, enables the isolation of subpopulations with different functional properties, isolation of transgenic cell lines, and purification of pluripotent cells from a heterogeneous cell population. However, it is important to confirm that current methods of sorting do not compromise the integrity or functional properties of these stem cells.
Viability of H9 hESCs following sorting
In the experiment shown, pluripotent H9 embryonic stem cells (TRA-1-81+, SSEA-3+, SSEA-1-) were sorted from a cultured population using a new comprehensive flow cytometry kit from BD Biosciences, the BD Stemflow™ human pluripotent stem cell sorting and analysis kit (Cat. No. 560461) on a BD FACSAria™ II cell sorter.
A) The plot shows the scatter plot of the cells established using BD™ CompBead Plus microparticles. Populations are shown as clusters defined by two markers.
Viability of H9 hESCs following sorting
In the experiment shown, pluripotent H9 embryonic stem cells (TRA-1-81+, SSEA-3+, SSEA-1-) were sorted from a cultured population using a new comprehensive flow cytometry kit from BD Biosciences, the BD Stemflow™ human pluripotent stem cell sorting and analysis kit (Cat. No. 560461) on a BD FACSAria™ II cell sorter.
B) To demonstrate cell viability, pluripotent cells were cultured following sorting, brightfield imaged, and examined for karyotypic abnormalities. Sorted cells were viable and displayed a normal karyotype.
Viability of H9 hESCs following sorting
In the experiment shown, pluripotent H9 embryonic stem cells (TRA-1-81+, SSEA-3+, SSEA-1-) were sorted from a cultured population using a new comprehensive flow cytometry kit from BD Biosciences, the BD Stemflow™ human pluripotent stem cell sorting and analysis kit (Cat. No. 560461) on a BD FACSAria™ II cell sorter.
C) To demonstrate cell viability, pluripotent cells were cultured following sorting, brightfield imaged, and examined for karyotypic abnormalities. Sorted cells were viable and displayed a normal karyotype.
Pluripotency of H9 hESCs following sorting
In the experiment shown, pluripotent H9 embryonic stem cells (TRA-1-81+, SSEA-3+, SSEA-1-) were sorted from a cultured population using the BD Stemflow™ Human Pluripotent Stem Cell Sorting and Analysis Kit (Cat. No. 560461) on a BD FACSAria™ II cell sorter.
A) Key markers for pluripotency (SSEA-4 Alexa Fluor® 647 (Cat. No. 560219) pseudocolored green, Oct3/4 Alexa Fluor® 555 (Cat. No. 560306) pseudocolored yellow, and SSEA-1 Alexa Fluor® 488 (Cat. No. 560172) pseudocolored red) were analyzed by imaging on a BD Pathway™ bioimager.
Pluripotency of H9 hESCs following sorting
In the experiment shown, pluripotent H9 embryonic stem cells (TRA-1-81+, SSEA-3+, SSEA-1-) were sorted from a cultured population using the BD™ Human Pluripotent Stem Celll Sorting and Analysis Kit (Cat. No. 560461) on a BD FACSAria™ II cell sorter.
B) Key markers for pluripotency (SSEA-4 Alexa Fluor® 647 (Cat. No. 560219) pseudocolored green, Oct 3/4 Alexa Fluor® 555 (Cat. No. 560306) pseudocolored yellow, and SSEA-1 Alexa Fluor® 488 (Cat. No. 560172) pseudocolored red) were analyzed by flow cytometry using the BD™ Human and Mouse Pluripotent Stem Cell Analysis Kit (Cat. No. 560477). Sorted cells expressed markers for pluripotency. Undifferentiated Oct3/4+SSEA-4+, SSEA-4+SSEA-1– cells composed 93.8% and 95.3% of the cell population, respectively.
