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The BD™ Human and Mouse Pluripotent Stem Cell Analysis Kit provides
a comprehensive research system for the reliable, indepth characterization
of cellular pluripotency and differentiation state in heterogeneous
stem cell mixtures.
To maximize reproducibility and improve productivity, the total
solution kit integrates pre-conjugated antibodies to markers for
pluripotency and differentiation (including the intracellular marker
Oct 3/4), compensation beads, staining and fixation buffers, verified
protocols, and software analysis guidelines. To provide the flexibility
to meet specific research objectives, an open design allows for
the easy addition of supplementary antibodies and the use of cell
lines expressing green fluorescent protein.
The differentiation status of human ES (H9) cells
was analyzed for multiple markers of pluripotency and differentiation
using the BD Human and Mouse Pluripotent Stem Cell Analysis Kit.
Undifferentiated Oct 3/4+SSEA-1- and Oct 3/4+SSEA-4+ human ES cells composed
94.9% and 98.7% of the cell population, respectively (panels
A and B). Histograms displaying cell counts versus the relative
expression level of individual markers are also shown (panels C, D, and
E).
Multicolor Flow Cytometry for In-depth Analysis
Capitalizing on flow cytometry’s powerful capabilities for
multiparameter analysis at the single-cell level, the system offers
researchers deep insight into cell identity and function. Data
on
the relative expression level of multiple markers can be obtained
for individual pluripotent or differentiated cells. Additionally,
the
kit offers access to data on absolute cell counts for particular
cell
subpopulations.
A Total Solution System to Minimize Variability
Monoclonal antibodies specific to two well known human
and mouse pluripotency markers* (Oct 3/4 PerCP-Cy™ 5.5
and SSEA-4 Alexa Fluor® 647) and one differentiation marker
(SSEA-1 PE) are pre-titrated and pre-conjugated to improve
productivity and reduce assay-to-assay variability. Corresponding
isotype controls are also included.
BD™ CompBead Plus microparticles simplify and standardize
experimental setup by facilitating compensation for multicolor
analysis. The use of BD CompBead Plus also significantly reduces
the number of cells required for experiments and controls.
Fixation and staining buffers, as well as protocols for fixation,
staining, and software analysis, standardize procedures and
reduce day-to-day and lab-to-lab variability.
Modular and Open to Accommodate Specific Needs
For simple customization and more advanced analysis, the open,
modular architecture of the kit allows for the easy addition of
supplementary monoclonal antibodies against critical cell-surface
markers. In addition, the kit uses a multicolor fluorochrome
combination that is compatible with cell lines expressing GFP.
BD CompBead Plus microparticles can also be used as compensation
controls for additional antibody drop-ins to this kit. This is
particularly useful when testing for markers that might or might
not be expressed on cells of interest.
A Resource for Stem Cell Research
With more than 20 years of successful experience in the field, BD Biosciences
continues to support innovation in the area of stem cell research.
Inspired by in-depth understanding of the complexities
of biological experiments, the BD Human and Mouse Pluripotent Stem Cell Analysis Kit
is designed to make it
easier for researchers to obtain accurate results, increase research
productivity, and accelerate discoveries.
*Note: When used with human stem cells, SSEA-4 is used as a pluripotency
marker and SSEA-1 as a differentiation marker. On mouse stem cells,
SSEA-4 can be used as a differentiation marker and SSEA-1 as a
mouse cell pluripotency marker.
The differentiation status of mouse ES (ES-E14TG2a) cells was
analyzed for multiple markers of pluripotency and differentiation
using the BD Human and Mouse Pluripotent Stem Cell Analysis Kit.
The markers for differentiation status in mouse are reversed from the human
markers. Undifferentiated Oct 3/4+SSEA-1+ and Oct 3/4+SSEA-4- cells composed
94.6% and 89.7% of the cell population, respectively (panels A and B). Histograms
displaying cell counts versus the relative expression level of individual
markers are also shown (panels C, D, and E). |
