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. 2022 Nov 25;11(23):3764. doi: 10.3390/cells11233764

Figure 1.

Figure 1

Molecular characterization of neuro–cardiac OOC composed of PC12 cells and hiPSC-CMs. (A) PC12 cells differentiate into parasympathetic neurons. PC12 were cultured for 14 days in the presence of 50 ng/mL NGF, fixed and immunolabelled for sympathetic markers. Representative immunofluorescence microscopy images show choline acetyltransferase (left panel; in red), tyrosine hydroxylase (central panel; in green), dopamine β-hydroxylase (right panel; in green), and β3-tubulin (right panel; in red). (B) Quantification of neurotransmitters in supernatant of PC12 cells. PC12 cells were differentiated for 14 days and subjected to addition of 1.5 mM Carbachol for 2 min. Then, medium was collected and the indicated neurotransmitters were quantified as described in the Materials and Methods section. (C) hiPSCs differentiate in cardiomyocytes (hiPSC-CMs) after 30 days following the so-called “matrix sandwich method” of differentiation. hiPSC-CMs were fixed and immunolabelled for sarcomeric cardiac markers. Representative immunofluorescence microscopy images show Cardiac troponin (in green) and the α-sarcomeric actinin (in red). Scale bars: 20 µm.