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(B) The representative phase contrast images were acquired (10x) at different time points during the p0 to p1 growth phase: d3, d5, and d7 post-plating.
A standard proliferation assay was utilized and viable cell numbers were quantified at 2, 7, 10, 14, and 21 days post-plating.
We next measured glucose uptake and lactate production in iPSC-derived cardiomyocytes 13 days post-plating, treated with trastuzumab up to 96 h.
Cells were microinjected at day 1 post-plating using a Cellinjector CI-2000 system (Fujitsu limited, Kanagawa, Japan) with 0.5 ± 0.2 μm injection pipettes (Femtotips, Eppendorf, NSW).
At 90 min or 5 h post-plating, slides were processed for immunostaining.
24 h post-plating, the media was removed and the cells rinsed with HBSS.
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The cells retain morphologic and biochemical characteristics of differentiated hepatocytes through day 30 post plating, including liver-specific gene expression.
At day 8 post plating, neuroepithelial clusters were also negative for CD73, however, ∼13% of the migrating emNCSCs were positive for CD73 (Figure 10B C).
We examined CD73 expression in day 5 neurospheres prior to culturing on fibronectin, as well as in epithelial clusters and migratory cells 3 days post plating on fibronectin.
The number of spheres was evaluated 6 days post plating.
Clonogenic survival was performed using post plating method of macro colony assay.
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