To ensure that iPSCs have been successfully reprogrammed and possess the properties of pluripotent stem cells, several characterization methods are employed:
Morphology: iPSCs are examined for their characteristic morphology, which includes a high nucleus-to-cytoplasm ratio and prominent nucleoli. Marker Expression: iPSCs are tested for the expression of pluripotency markers such as Oct4, Nanog, and SSEA-4 through techniques like immunocytochemistry and flow cytometry. Differentiation Potential: The ability of iPSCs to differentiate into all three germ layers (ectoderm, mesoderm, and endoderm) is assessed through directed differentiation assays and teratoma formation. Genetic Integrity: Genomic stability is checked using karyotyping and next-generation sequencing to detect any chromosomal abnormalities or mutations.
