Method Profile
Four donor-derived cardiac cell lines support parallel GPCR signaling, calcium, extracellular electrical and impedance measurements.
Explore individual pharmacological response profiles and test how molecular signaling relates to functional cardiac-cell behavior.
One induced pluripotent stem cell (iPSC)-derived cardiomyocyte line from each of two healthy controls and two patients with nonischemic dilated cardiomyopathy.
Nuclear/cytoplasmic PKA and ERK biosensors, spontaneous calcium imaging and separate extracellular field-potential/1-Hz-paced impedance measurements before and after ligands.
Human cells
Four blood-derived iPSC lines validated for pluripotency, karyotype and cardiomyocyte identity; approximately 85% cardiomyocyte purity with the reported differentiation.
Pharmacological probes
Angiotensin, adrenergic and endothelin-receptor ligands; key functional comparisons include 10 μM isoproterenol, phenylephrine and endothelin-1 dose responses.
Measurements
Compartment-targeted PKA/ERK reporters, myofilament-localized RGECO-TnT calcium reporter, extracellular field potentials and impedance.
Compare each donor’s signaling and functional endpoints separately, retaining the distinction between spontaneously measured calcium events and 1-Hz-paced impedance; impedance amplitude is not absolute contractile force.
Compare baseline and ligand-stimulated molecular/function profiles across the four donor cell lines.
Two control donors and two DCM patients, one line per donor; signaling plates, cells and recording wells provide assay-level replication.
Matched vehicle or pretreatment baseline within each line and the corresponding other donor lines.
One patient line showed elevated phenylephrine-associated cytosolic PKA without a corresponding dose-dependent beat-rate/impedance increase. A second patient line showed little isoproterenol frequency response in calcium imaging but an increase in field-potential recordings.
A four-donor study compares nuclear and cytoplasmic signaling with calcium, electrical and impedance measurements. Distinct responses in DCM-derived cells show why increased signaling cannot alone establish improved cardiac-cell function.
