<> "The repository administrator has not yet configured an RDF license."^^ . <> . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells"^^ . "Background: Human induced pluripotent stem cells (hiPSC) harbor the potential to differentiate into diverse cardiac cell types. Previous experimental efforts were primarily directed at the generation of hiPSC-derived cells with ventricular cardiomyocyte characteristics. Aiming at a straightforward approach for pacemaker cell modeling and replacement, we sought to selectively differentiate cells with nodal-type properties.\r\n\r\nMethods: hiPSC were differentiated into spontaneously beating clusters by co-culturing with visceral endoderm-like cells in a serum-free medium. Subsequent culturing in a specified fetal bovine serum (FBS)-enriched cell medium produced a pacemaker-type phenotype that was studied in detail using quantitative real-time polymerase chain reaction (qRT-PCR), immunocytochemistry, and patch-clamp electrophysiology. Further investigations comprised pharmacological stimulations and co-culturing with neonatal cardiomyocytes.\r\n\r\nResults: hiPSC co-cultured in a serum-free medium with the visceral endoderm-like cell line END-2 produced spontaneously beating clusters after 10–12 days of culture. The pacemaker-specific genes HCN4, TBX3, and TBX18 were abundantly expressed at this early developmental stage, while levels of sarcomeric gene products remained low. We observed that working-type cardiomyogenic differentiation can be suppressed by transfer of early clusters into a FBS-enriched cell medium immediately after beating onset. After 6 weeks under these conditions, sinoatrial node (SAN) hallmark genes remained at high levels, while working-type myocardial transcripts (NKX2.5, TBX5) were low. Clusters were characterized by regular activity and robust beating rates (70–90 beats/min) and were triggered by spontaneous Ca2+ transients recapitulating calcium clock properties of genuine pacemaker cells. They were responsive to adrenergic/cholinergic stimulation and able to pace neonatal rat ventricular myocytes in co-culture experiments. Action potential (AP) measurements of cells individualized from clusters exhibited nodal-type (63.4%) and atrial-type (36.6%) AP morphologies, while ventricular AP configurations were not observed.\r\n\r\nConclusion: We provide a novel culture media-based, transgene-free approach for targeted generation of hiPSC-derived pacemaker-type cells that grow in clusters and offer the potential for disease modeling, drug testing, and individualized cell-based replacement therapy of the SAN."^^ . "2017" . . "8" . "229" . . "BioMed Central"^^ . . . "Stem Cell Research & Therapy"^^ . . . "17576512" . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . "Dierk"^^ . "Thomas"^^ . "Dierk Thomas"^^ . . "Claudia"^^ . "Seyler"^^ . "Claudia Seyler"^^ . . "Hugo A."^^ . "Katus"^^ . "Hugo A. Katus"^^ . . "Nina D."^^ . "Ullrich"^^ . "Nina D. Ullrich"^^ . . "Pascal"^^ . "Geschwill"^^ . "Pascal Geschwill"^^ . . "Andreas"^^ . "Draguhn"^^ . "Andreas Draguhn"^^ . . "Rasmus"^^ . "Rivinius"^^ . "Rasmus Rivinius"^^ . . "Boris"^^ . "Greber"^^ . "Boris Greber"^^ . . "Jochen"^^ . "Utikal"^^ . "Jochen Utikal"^^ . . "Fabrice F."^^ . "Darche"^^ . "Fabrice F. Darche"^^ . . "Karin"^^ . "Müller-Decker"^^ . "Karin Müller-Decker"^^ . . "Michael"^^ . "Koenen"^^ . "Michael Koenen"^^ . . "Patrick A."^^ . "Schweizer"^^ . "Patrick A. Schweizer"^^ . . . . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells (PDF)"^^ . . . "13287_2017_Article_681.pdf"^^ . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells (Other)"^^ . . . . . . "preview.jpg"^^ . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells (Other)"^^ . . . . . . "medium.jpg"^^ . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells (Other)"^^ . . . . . . "small.jpg"^^ . . . "Subtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells (Other)"^^ . . . . . . "indexcodes.txt"^^ . . "HTML Summary of #23646 \n\nSubtype-specific differentiation of cardiac pacemaker cell clusters from human induced pluripotent stem cells\n\n" . "text/html" . . . "610 Medizin"@de . "610 Medical sciences Medicine"@en . .