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The oxidation of FG in acidic oxidizing medium can be interpreted as a contracting area process controlled by phase boundary.
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Using the hanging drop technique and induction with ascorbate, both individual and multiple foci of spontaneously contracting areas were observed within one day of EB plating.
For example, we have found that, even in the absence of specific chemical or molecular manipulation, the frequency of formation of spontaneously contracting areas could vary substantially, depending on FBS concentration and density of EBs plated per well as well as stochastic events.
Spontaneously contracting areas were manually dissected at day 20 30 of differentiation.
Spontaneously contracting areas were manually dissected and cultured further until days 20 60 of differentiation.
Control- and CPVT-iPSC lines developed spontaneously contracting areas (Supplementary Movie 1) expressing cardiac-specific channel and structural genes.
Spontaneously contracting areas, which appeared 12 30 days after EB plating, were manually microdissected and plated onto fibronectin-coated plates for further differentiation for an additional 45 90 days.
EB differentiation was achieved by aggregating the cells on low-attachment plates and EBs were plated on gelatin-coated dishes at day 7. Spontaneously contracting areas were manually dissected and cultured further until day 90 130 of differentiation.
The frequency of antral contractions was defined as the number of contractions during a 25-minute interval, recorded per five minutes, and the antral contractions were defined as one time while the maximal contraction-induced reduction of the antral area (difference between relaxed and contracted area) as a fraction of the relaxed area (Δ A/ A) > 50% as our previous study [ 11].
The contemporary genetic sectors in Iberian F. ceranoides are more likely to result from the regional reassembly of vicariant phylogroups into their current distributions following a period of independent, mostly allopatric divergence in past contracted areas of occurrence.
The EBs were plated after 10 days on gelatin-coated culture dishes and observed for the appearance of spontaneous contracting areas.
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