Introduction of Karyotype Analysis for iPSC
The technology of induced pluripotent stem cell (iPSC) lines generation provides a valuable resource for disease modeling, regenerative therapy, and development study. However, the accumulation of karyotypic during long-term iPSCs culture results in chromosomal abnormalities, changes in gene expression and cellular functions, and even increases the risk of the iPSCs being tumorigenic. As genomic alterations present potential risks in the iPSCs applications, it is important to monitor the genomic integrity of these iPSCs lines especially in iPSCs intended for therapeutic use. Now Creative Biolabs has developed various strategies for the iPSC karyotype analysis. iPSC karyotype analysis is the examination of morphology. Moreover, the changes in size, the position of centromeres, and banding patterns are also necessary for detection aspects.
G-bonding Karyotyping for iPSC
The typical strategies for iPSC karyotype analysis are based on staining the chromosomes of metaphase cells in distinct banding patterns. G-banding is a technique to produce a visible karyotype by staining condensed chromosomes. After partially digesting with trypsin, the metaphase chromosomes are stained with Giemsa stain. Heterochromatic regions is more darkly stained in G-banding while less condensed chromatin present light bands in G-banding. We always recommend that a stem cell line should be karyotyped every 10–15 passages to ensure that there are no chromosomal duplications, insertions, deletions, translocations, or centromere loss during the long-term culture. Moreover, we can also provide various karyotyping methods including C-banding (constitutive heterochromatin staining), R-banding (reverse Giemsa staining), T-banding (telomeric staining), and Q-banding (quinacrine staining). In summary, all these methods are based on the abnormality identification according to the changes in banding patterns.
