Karyotyping analysis of H9 hESCs was conducted using standard G-banding techniques. The hESCs were treated with 0.1 mg/mL of colcemid KaryoMAX for 3 h. After centrifuging the cells, the cell pellets were resuspended in prewarmed 1% of sodium citrate solution and incubated for 50 min. The cells were pelleted by centrifugation and resuspended in fixative solution (3:1 (v/v) methanol/acetic acid). Metaphase spreads were prepared on glass microscope slides and G-banded by exposure to trypsin and stained with 3:1 Gurr’s/Leishmann’s stain.

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