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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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In a surprise move this May, China became the first country to apply for a contract area in international waters to prospect for massive sulphide ore.
The EBs were plated after 10 days on gelatin-coated culture dishes and observed for the appearance of spontaneous contracting areas.
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.
Substantial increases in contracting areas were also noted with dorsomorphin treatment of unmodified CGR8 and R1 mouse ES cell lines maintained by three independent laboratories.
To confirm that the large synchronously contracting areas of DsRed+ cells induced by the dorsomorphin treatment were composed of cardiomyocytes, dorsomorphin EBs were fixed at day 12 and processed for immunostaining with specific antibodies against several known cardiac markers.
Although hESC differentiation efficiencies up to 70% (as assessed by the percentage of contracting hEB generated) have been published [3], the most commonly used basic protocol for hESC cardiac differentiation has a low efficiency of ∼8 22% [6], [7], and takes up to 21 days to produce contracting areas.
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.
Immunostaining analysis confirmed the presence and the distribution of RyR2 in cells isolated from the contracting areas.
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