Mouse embryonic stem cells (mESCs) are unique cells in that they can be maintained in vitro as pluripotent cells and can be induced to differentiate into many cell types by manipulating culture conditions. mESCs have been used as a model to help define differentiation and signaling pathways in human embryonic stem cells (hESCs), since hESCs are more arduous to work on. Therefore, much attention has been focused on understanding the mechanisms behind mESC differentiation.
Currently, the most utilized method to test the differentiation state of mESCs is to analyze the expression of the proteins OCT4 and the stage specific embryonic antigen, SSEA1 (1,2). The transcription factor, OCT4, is highly expressed in undifferentiated mESCs and is essential for the formation and maintenance of pluripotentcy (3,4). SSEA1 is also expressed on undiiferentiated mESCs but another stage specific embryonic antigen, SSEA4, is not (2,5,6). However, upon retinoic treatment, SSEA1 expression wanes and in some instances, accompanied by expression of SSEA4 (6,7).
The FlowCellect™ Mouse Embryonic Stem Cell Characterization Kit uses an antibody-based detection method, utilizing fluorescently conjugated antibodies to detect OCT4, SSEA4 and SSEA1. The FlowCellect™ Mouse Embryonic Stem Cell Characterization kit is designed specifically for flow cytometers with a single blue (488 nm) laser light source. The FlowCellect™ Mouse Embryonic Stem Cell Characterization Kit is not intended for use in microscopy applications including ICC, IHC and HTS, western blot or any other fluorometric analysis platforms.
The kit is optimized for use on the Guava EasyCyte™ Plus cytometer that requires cell concentrations to be less than 500 cells per microliter. The kit does not include culture media or other materials that should be readily available in a common laboratory setting that routinely performs experiments of the type described here.