Description
• Measure the ubiquitous alkaline phosphatase marker in embryonic stem cells and embryonic germ cells
• Quantify activity by colorimetric or fluorometric plate reader
Alkaline Phosphatase (AP) is a widely used marker for both mouse and human embryonic stem cells (ES) and embryonic germ cells (EG). Our StemTAG Alkaline Phosphatase kits provide an efficient system for monitoring cell differentiation or undifferentiation using the AP marker. The StemTAG Alkaline Phosphatase Activity Assay Kits provide reagents for quantifying alkaline phosphatase activity in a convenient 96-well plate format, with either colorimetric or fluorescence detection.
Introduction: Embryonic stem (ES) cells are continuous proliferating stem cell lines of embryonic origin first isolated from the inner cell mass (ICM). Two distinguishing features of ES cells are their ability to be maintained indefinitely in an undifferentiated state and their potential to develop into any cell within the body. Based on previous methods developed for mouse ES cells, human ES cell lines were first established by Dr. James Thomson and colleagues. Like mouse ES cells, human ES cells express high levels of membrane alkaline phosphatase (AP) and Oct-4, a transcriptional factor critical to ICM and germline formation. However, unlike mouse ES cells, hES cells do not express stage-specific embryonic antigen (SSEA-1). In addition, prolonged propagation of hES cells is typically achieved by coculture with primary mouse embryonic fibroblasts (MEFs) serving as feeder cells. Human ES cell lines are not able to maintain their undifferentiated state in the absence of supporting feeder layer cells, even when exogenous cytokines such as leukemia inhibitory factor (LIF) and gelatin-coated plates are used.
Table 1. Comparison of Mouse and Human Pluripotent Stem Cells.
Although stem cells from different origins require different growth conditions for self-renewal and display different cell surface markers (see Table 1), AP is the most widely used stem cell marker. The StemTAG Alkaline Phosphatase Activity Assay Kit provides an efficient system for monitoring ES cell undifferentiation/ differentiation through AP activity by quantitative assay