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Comparative analysis of different passages revealed that methylation changes are continuously acquired with every passage upon long-term culture (Fig. S4B).
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Cells were maintained and passaged upon reaching confluence by standard cell culture techniques.
MSCs were passaged upon reaching 50% confluency and cells from passage 2 4 were used for this study.
Human fibroblasts were isolated from fetal liver and grown in Isocove's modified Dulbecco's medium containing 10% FBS, and passaged upon confluence [1].
SFFs were passaged upon reaching 95% confluence with 0.25% trypsin-EDTA and infected with reprogramming virus between passages 3 and 5. 293T cells (ATCC, Manassas, VA, USA) were maintained in the same medium.
Cells were passaged upon reaching confluence.
Cells were passaged upon 75-80% confluence and counted.
Generally, cells were passaged upon reaching 75% confluence.
These cells were passaged upon reaching confluence over a 6-month period.
Treatment media was replaced every 4 5 days, and cells were passaged upon reaching confluence.
Growth media was replaced every 3 days and cultures were passaged upon reaching 70-80% confluence.
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