Stem cell experiments often require weeks of culturing cells before a hypothesis can be tested. Stem cell products from R&D Systems provide cost-effective tools to verify the potency and health of starting cell populations, a quality control step that can reduce experimental variability and prevent weeks of lost time and reagents.
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Verification of Induced Pluripotent Stem Cell Pluripotency. iPS2 human induced pluripotent stem cells were differentiated to ectoderm, mesoderm, and endoderm using the media supplements included in the Human Pluripotent Stem Cell Functional Identification Kit (Catalog # SC027). The kit also contains Goat Anti-Human Otx2 (ectoderm), Goat Anti-Human Brachyury (mesoderm), and Goat Anti-Human SOX17 (endoderm) Antigen Affinity Purified Polyclonal Antibodies for the confirmation of differentiation status. To further evaluate lineage commitment, cells were stained with a Goat Anti-Human SOX1 Antigen Affinity Purified Antibody (Catalog # AF3369), a Goat Anti-Human HAND1 Antigen Affinity Purified Antibody (Catalog # AF3168), and a Goat Anti-Human HNF-3 beta/FoxA2 Antigen Affinity Purified Antibody (Catalog # AF2400). The cells were stained using the NorthernLights™ 557-conjugated Donkey Anti-Goat IgG Secondary Antibody (Catalog # NL001; red) and the nuclei were counterstained with DAPI (blue). .
Functional Identification Kits Can Be Used Alone or in Combination with Our Other Stem Cell Kits. See Example Data:
Verification of Germ Layer Markers in Differentiated Pluripotent Stem Cells. iPS2 human induced pluripotent stem cells were differentiated into each of the three germ layers using the base media and differentiation supplements provided in the Human Pluripotent Stem Cell Functional Identification Kit (Catalog # SC027). Germ layer differentiation was subsequently verified using the six fluorochrome-conjugated antibodies provided in the Human Three Germ Layer 3-Color Immunocytochemistry Kit (Catalog # SC022). Ectoderm differentiated cells were simultaneously stained with NorthernLights (NL) fluorochrome NL557-conjugated Otx2 (red) and NL493-conjugated SOX1 (green). Mesoderm differentiated cells were simultaneously stained with NL557-conjugated Brachyury (red) and NL637-conjugated HAND1 (green). Endoderm differentiated cells were simultaneously stained with NL637-conjugated SOX17 (red) and NL493-conjugated GATA-4 (green). All nuclei were counterstained with DAPI (blue). Nuclear expression of each marker was detected in its respective cell lineage.