TY - JOUR AU - Németh, Krisztina AU - László, Zsófia AU - Biró, Adrienn AU - Szatmári, Ágnes AU - Cserép, Balázs Gergely AU - Várady, György AU - Bakos, Éva AU - Laczka, Csilla AU - Kele, Péter TI - Organic anion transporting polypeptide 3A1 (OATP3A1)-gated bioorthogonal labeling of intracellular proteins JF - MOLECULES J2 - MOLECULES VL - 28 PY - 2023 IS - 6 PG - 10 SN - 1420-3049 DO - 10.3390/molecules28062521 UR - https://m2.mtmt.hu/api/publication/33688209 ID - 33688209 N1 - Chemical Biology Research Group, Institute of Organic Chemistry, RCNS, Magyar Tudósok Krt. 2, Budapest, H-1117, Hungary Molecular Cell Biology Research Group, Institute of Enzymology, RCNS, Magyar Tudósok Krt. 2, Budapest, H-1117, Hungary Membrane Protein Research Group, Institute of Enzymology, RCNS, Magyar Tudósok Krt. 2, Budapest, H-1117, Hungary Export Date: 19 April 2023 CODEN: MOLEF Correspondence Address: Németh, K.; Chemical Biology Research Group, Magyar Tudósok Krt. 2, Hungary; email: nemeth.krisztina@ttk.hu Correspondence Address: Kele, P.; Chemical Biology Research Group, Magyar Tudósok Krt. 2, Hungary; email: kele.peter@ttk.hu Chemicals/CAS: protein, 67254-75-5; Fluorescent Dyes; Organic Anion Transporters; Proteins Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFIH, NKFIH-K-138518, NKFIH-K-143581, VEKOP-2.3.3-15-2016-00011 Funding text 1: Present work was supported by the National Research, Development and Innovation Office of Hungary (NKFIH-K-143581, NKFIH-K-138518 and VEKOP-2.3.3-15-2016-00011). AB - Organic anion transporting polypeptides (OATPs) were found to readily deliver membrane impermeable, tetrazine bearing fluorescent probes into cells. This feature was explored in OATP3A1 conditioned bio-orthogonal labeling schemes of various intracellular proteins in live cells. Confocal microscopy and super-resolution microscopy (STED) studies have shown that highly specific and efficient staining of the selected intracellular proteins can be achieved with the otherwise non-permeable probes when OATP3A1 is present in the cell membrane of cells. Such a transport protein linked bio-orthogonal labeling scheme is believed to be useful in OATP3A1 activity-controlled protein expression studies in the future. © 2023 by the authors. LA - English DB - MTMT ER - TY - JOUR AU - Szatmári, Ágnes AU - Cserép, Balázs Gergely AU - Molnár , Tibor Ákos AU - Söveges, Bianka AU - Biró, Adrienn AU - Várady, György AU - Szabó, Edit Zsuzsanna AU - Németh, Krisztina AU - Kele, Péter TI - A Genetically Encoded Isonitrile Lysine for Orthogonal Bioorthogonal Labeling Schemes JF - MOLECULES J2 - MOLECULES VL - 26 PY - 2021 IS - 16 PG - 16 SN - 1420-3049 DO - 10.3390/molecules26164988 UR - https://m2.mtmt.hu/api/publication/32172729 ID - 32172729 N1 - Chemical Biology Research Group, Institute of Organic Chemistry, ELKH Research Centre for Natural Sciences, Magyar Tudósok Krt 2, Budapest, H-1117, Hungary Molecular Cell Biology Research Group, Institute of Enzymology, ELKH Research Centre for Natural Sciences, Magyar Tudósok Krt 2, Budapest, H-1117, Hungary Cited By :6 Export Date: 31 October 2023 CODEN: MOLEF Correspondence Address: Szatmári, Á.; Chemical Biology Research Group, Magyar Tudósok Krt 2, Hungary; email: szatmari.agnes@ttk.hu Correspondence Address: Németh, K.; Chemical Biology Research Group, Magyar Tudósok Krt 2, Hungary; email: nemeth.krisztina@ttk.hu Correspondence Address: Kele, P.; Chemical Biology Research Group, Magyar Tudósok Krt 2, Hungary; email: kele.peter@ttk.hu LA - English DB - MTMT ER - TY - JOUR AU - Kovács, Réka AU - Vadászi, Henrietta AU - Bulyáki, Éva AU - Török, György AU - Tóth, Vilmos AU - Mátyás, Dominik AU - Kun, Judit AU - Hunyadi-Gulyás Éva, Csilla AU - Fedor, Flóra Zsófia AU - Csincsi, Ádám AU - Medzihradszky F., Katalin AU - Homolya, László AU - Juhász, Gábor Dénes AU - Kékesi, Adrienna Katalin AU - Józsi, Mihály AU - Györffy, Balázs A. AU - Kardos, József TI - Identification of Neuronal Pentraxins as Synaptic Binding Partners of C1q and the Involvement of NP1 in Synaptic Pruning in Adult Mice JF - FRONTIERS IN IMMUNOLOGY J2 - FRONT IMMUNOL VL - 11 PY - 2021 PG - 17 SN - 1664-3224 DO - 10.3389/fimmu.2020.599771 UR - https://m2.mtmt.hu/api/publication/31861935 ID - 31861935 N1 - ELTE NAP Neuroimmunology Research Group, Department of Biochemistry, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Molecular Cell Biology Research Group, Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Sciences Centre of Excellence, Budapest, Hungary Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary Laboratory of Proteomics, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Laboratory of Proteomics Research, Biological Research Centre, Eötvös Loránd Research Network (ELKH), Szeged, Hungary Doctoral School of Chemical Engineering and Material Sciences, Pannon University, Veszprém, Hungary Complement Research Group, Department of Immunology, ELTE Eötvös Loránd University, Budapest, Hungary Department of Physiology and Neurobiology, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Cited By :4 Export Date: 23 July 2022 Correspondence Address: Kardos, J.; ELTE NAP Neuroimmunology Research Group, Hungary LA - English DB - MTMT ER - TY - JOUR AU - Török, György AU - Cserép, Balázs Gergely AU - Telek, András AU - Arany, Dóra AU - Váradi, Melinda AU - Homolya, László AU - Kellermayer, Miklós AU - Kele, Péter AU - Németh, Krisztina TI - Large Stokes-shift bioorthogonal probes for STED, 2P-STED and multi-color STED nanoscopy JF - METHODS AND APPLICATIONS IN FLUORESCENCE J2 - METHODS APPL FLUORESC VL - 9 PY - 2021 IS - 1 PG - 13 SN - 2050-6120 DO - 10.1088/2050-6120/abb363 UR - https://m2.mtmt.hu/api/publication/31523926 ID - 31523926 LA - English DB - MTMT ER - TY - JOUR AU - Hathy, Edit Margit AU - Szabó, Eszter AU - Varga, Nóra AU - Erdei, Zsuzsa AU - Tordai, Csongor AU - Czehlár, Boróka AU - Baradits, Máté AU - Jezsó, Bálint AU - Koller, Júlia AU - Nagy, László AU - Molnár, Mária Judit AU - Homolya, László AU - Nemoda, Zsófia AU - Apáti, Ágota AU - Réthelyi, János TI - Investigation of de novo mutations in a schizophrenia case-parent trio by induced pluripotent stem cell-based in vitro disease modeling: convergence of schizophrenia- and autism-related cellular phenotypes JF - STEM CELL RESEARCH & THERAPY J2 - STEM CELL RES THER VL - 11 PY - 2020 IS - 1 PG - 15 SN - 1757-6512 DO - 10.1186/s13287-020-01980-5 UR - https://m2.mtmt.hu/api/publication/31678126 ID - 31678126 N1 - National Brain Research Project (NAP) Molecular Psychiatry Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest, Hungary Molecular Cell Biology Research Group, Institute of Enzymology, Research Center for Natural Sciences, 1117 Magyar tudósok körútja 2, Budapest, Hungary Institute of Rare Disorders and Genomic Medicine, Semmelweis University, Budapest, Hungary Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, Budapest, Hungary Department of Psychiatry and Psychotherapy, Semmelweis University, Balassa utca 6, Budapest, 1083, Hungary Cited By :6 Export Date: 18 November 2022 Correspondence Address: Réthelyi, J.M.; Department of Psychiatry and Psychotherapy, Balassa utca 6, Hungary; email: rethelyi.janos@med.semmelweis-univ.hu Correspondence Address: Apáti, Á.; Molecular Cell Biology Research Group, 1117 Magyar tudósok körútja 2, Hungary; email: apati.agota@ttk.hu LA - English DB - MTMT ER - TY - JOUR AU - Györffy, Balázs AU - Tóth, Vilmos AU - Török, György AU - Gulyássy, P. AU - Kovács, Réka AU - Vadászi, Henrietta AU - Micsonai, András AU - Tóth, Erzsébet Melinda AU - Sántha, Miklós AU - Homolya, László AU - Drahos, László AU - Juhász, Gábor Dénes AU - Kékesi, Adrienna Katalin AU - Kardos, József TI - Synaptic mitochondrial dysfunction and septin accumulation are linked to complement-mediated synapse loss in an Alzheimer’s disease animal model JF - CELLULAR AND MOLECULAR LIFE SCIENCES J2 - CELL MOL LIFE SCI VL - 77 PY - 2020 IS - 24 SP - 5243 EP - 5258 PG - 16 SN - 1420-682X DO - 10.1007/s00018-020-03468-0 UR - https://m2.mtmt.hu/api/publication/31177389 ID - 31177389 N1 - ELTE NAP Neuroimmunology Research Group, Department of Biochemistry, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Laboratory of Proteomics, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Department of Biochemistry, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Molecular Cell Biology Research Group, Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences Centre of Excellence, Budapest, Hungary Department of Biophysics and Radiation Biology, Semmelweis University, Budapest, Hungary MS Proteomics Research Group, Institute of Organic Chemistry, Research Centre for Natural Sciences, Budapest, Hungary Institute of Biochemistry, Biological Research Centre, Szeged, Hungary CRU Hungary Ltd., Göd, Hungary Department of Physiology and Neurobiology, Institute of Biology, ELTE Eötvös Loránd University, Budapest, Hungary Export Date: 14 February 2020 CODEN: CMLSF Correspondence Address: Kardos, J.; ELTE NAP Neuroimmunology Research Group, Department of Biochemistry, Institute of Biology, ELTE Eötvös Loránd UniversityHungary; email: kardos@elte.hu Funding details: KTIA_NAP_13-2-2014-0017, K120391, FIEK_16-1-2016-0005, KTIA_NAP_13-2-2015-0003, OTKA–K128123 Funding details: Semmelweis Egyetem Funding details: Emberi Eroforrások Minisztériuma Funding details: Magyar Tudományos Akadémia, MTA Funding text 1: Open access funding provided by Eötvös Loránd University (ELTE). This study was supported by the National Research, Development and Innovation Office of Hungary Grants KTIA_NAP_13-2-2014-0017 and 2017-1.2.1-NKP-2017-00002 (to BAG, VT, GJ, AM, KAK, and JK), FIEK_16-1-2016-0005 (to BAG, RÁK, HV, AM, GJ, KAK, and JK), KTIA_NAP_13-2-2015-0003 (to PG, LD, and GJ), OTKA–K128123 (to LH), and K120391 (to JK and AM). The research was partly financed by the Higher Education Institutional Excellence Programme of the Ministry of Human Capacities in Hungary, within the framework of the Therapeutic Development thematic programme of the Semmelweis University (GT). AM is supported by the Bolyai János fellowship of the Hungarian Academy of Sciences. AB - Synaptic functional disturbances with concomitant synapse loss represent central pathological hallmarks of Alzheimer’s disease. Excessive accumulation of cytotoxic amyloid oligomers is widely recognized as a key event that underlies neurodegeneration. Certain complement components are crucial instruments of widespread synapse loss because they can tag synapses with functional impairments leading to their engulfment by microglia. However, an exact understanding of the affected synaptic functions that predispose to complement-mediated synapse elimination is lacking. Therefore, we conducted systematic proteomic examinations on synaptosomes prepared from an amyloidogenic mouse model of Alzheimer’s disease (APP/PS1). Synaptic fractions were separated according to the presence of the C1q-tag using fluorescence-activated synaptosome sorting and subjected to proteomic comparisons. The results raised the decline of mitochondrial functions in the C1q-tagged synapses of APP/PS1 mice based on enrichment analyses, which was verified using flow cytometry. Additionally, proteomics results revealed extensive alterations in the level of septin protein family members, which are known to dynamically form highly organized pre- and postsynaptic supramolecular structures, thereby affecting synaptic transmission. High-resolution microscopy investigations demonstrated that synapses with considerable amounts of septin-3 and septin-5 show increased accumulation of C1q in APP/PS1 mice compared to the wild-type ones. Moreover, a strong positive correlation was apparent between synaptic septin-3 levels and C1q deposition as revealed via flow cytometry and confocal microscopy examinations. In sum, our results imply that deterioration of synaptic mitochondrial functions and alterations in the organization of synaptic septins are associated with complement-dependent synapse loss in Alzheimer’s disease. © 2020, The Author(s). LA - English DB - MTMT ER - TY - JOUR AU - Török, György AU - Erdei, Zsuzsa AU - Lilienberg, Julianna AU - Apáti, Ágota AU - Homolya, László TI - The importance of transporters and cell polarization for the evaluation of human stem cell-derived hepatic cells JF - PLOS ONE J2 - PLOS ONE VL - 15 PY - 2020 IS - 1 PG - 22 SN - 1932-6203 DO - 10.1371/journal.pone.0227751 UR - https://m2.mtmt.hu/api/publication/31139755 ID - 31139755 AB - One of the most promising applications of human pluripotent stem cells is their utilization for human-based pharmacological models. Despite the fact that membrane transporters expressed in the liver play pivotal role in various hepatic functions, thus far only little attention was devoted to the membrane transporter composition of the stem cell-derived liver models. In the present work, we have differentiated HUES9, a human embryonic stem cell line, toward the hepatic lineage, and monitored the expression levels of numerous differentiation marker and liver transporter genes with special focus on ABC transporters. In addition, the effect of bile acid treatment and polarizing culturing conditions on hepatic maturation has been assessed. We found that most transporter genes crucial for hepatic functions are markedly induced during hepatic differentiation; however, as regards the transporter composition the end-stage cells still exhibited dual, hepatocyte and cholangiocyte character. Although the bile acid treatment and sandwich culturing only slightly influenced the gene expressions, the stimulated cell polarization resulted in formation of bile canaliculi and proper localization of transporters. Our results point to the importance of membrane transporters in human stem cell-derived hepatic models and demonstrate the relevance of cell polarization in generation of applicable cellular models with correctly localized transporters. On the basis of our observations we suggest that conventional criteria for the evaluation of the quality of stem cell-derived hepatocyte-like cells ought to be augmented with additional elements, such as polarized and functional expression of hepatic transporters. LA - English DB - MTMT ER - TY - JOUR AU - Bakos, Éva AU - Nemet, Orsolya AU - Patik, Izabel AU - Kucsma, Nóra AU - Várady, György AU - Szakács, Gergely AU - Laczka, Csilla TI - A novel fluorescence-based functional assay for human OATP1A2 and OATP1C1 identifies interaction between third-generation P-gp inhibitors and OATP1A2 JF - FEBS JOURNAL J2 - FEBS J VL - 287 PY - 2020 IS - 12 SP - 2468 EP - 2485 PG - 18 SN - 1742-464X DO - 10.1111/febs.15156 UR - https://m2.mtmt.hu/api/publication/31069992 ID - 31069992 N1 - Membrane Protein Research Group, Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary Laboratory of Molecular Cell Biology, Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary Institute of Cancer Research, Medical University Vienna, Wien, Austria Cited By :1 Export Date: 22 July 2020 CODEN: FJEOA Correspondence Address: Özvegy-Laczka, C.; Membrane Protein Research Group, Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of SciencesHungary; email: laczka.csilla@ttk.mta.hu Chemicals/CAS: elacridar, 143664-11-3; levofloxacin, 100986-85-4, 138199-71-0; methotrexate, 15475-56-6, 59-05-2, 7413-34-5; quinine, 130-89-2, 130-95-0, 14358-44-2, 549-48-4, 549-49-5, 60-93-5, 7549-43-1; tariquidar, 206873-63-4; verapamil, 152-11-4, 52-53-9; zosuquidar, 167354-41-8, 167465-36-3, 312905-17-2, 474276-97-6 Tradenames: gf120918 Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA, FK 128751 Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFI Funding text 1: We appreciate the financial support from the National Research Development and Innovation Office (OTKA, grant number FK 128751). CÖ‐L is also grateful for the support by the János Bolyai Fellowship of the Hungarian Academy of Sciences. Membrane Protein Research Group, Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary Laboratory of Molecular Cell Biology, Institute of Enzymology, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary Institute of Cancer Research, Medical University Vienna, Wien, Austria Cited By :5 Export Date: 26 August 2021 CODEN: FJEOA Correspondence Address: Özvegy-Laczka, C.; Membrane Protein Research Group, Hungary; email: laczka.csilla@ttk.mta.hu AB - Organic anion-transporting polypeptide 1A2 (OATP1A2), expressed in the human blood-brain barrier, promotes drug uptake from the blood and hence can be exploited for central nervous system-targeted drug delivery. The thyroid transporter OATP1C1, expressed in the choroid plexus and in astrocytes, is also a potential pharmacological target. Based on their established pharmacological relevance, screening the drug interaction profile of OATP1A2 and OATP1C1 is highly desirable. However, drug interaction screens require suitable model systems and functional assays. In the current study, uptake of a set of cell-impermeable fluorescent dyes was screened in HEK-293 and A431 cell lines overexpressing OATP1A2 and OATP1C1. Based on the uptake of fluorescent dye substrates, a functional assay was developed, which was used to characterize OATP inhibitors/substrates. We identify Live/Dead Green (LDG), Live-or-Dye 488, and sulforhodamines 101, G, and B as novel fluorescent substrates of OATP1A2 and OATP1C1. We show that LDG uptake is proportional to OATP1A2/1C1 expression, allowing the isolation of cells expressing high transporter levels. Additionally, dye uptake can be used to characterize the drug interaction pattern of OATP1A2 and OATP1C1. We demonstrate that third-generation P-glycoprotein inhibitors elacridar, tariquidar, and zosuquidar inhibit OATP1A2 function. Increased toxicity of elacridar in OATP1A2-expressing cells suggests that OATP1A2 may modulate the distribution of this compound. The fluorescence-based assays developed in the current study are a good alternative of radioligand-based tests and pave the way toward high-throughput screens for OATP1A2/1C1 drug interaction studies. LA - English DB - MTMT ER - TY - JOUR AU - Györffy, Balázs AU - Kun, Judit AU - Török, György AU - Bulyáki, Éva AU - Borhegyi, Zsolt AU - Gulyássy, Péter AU - Kis, Viktor AU - Szocsics, Péter AU - Micsonai, András AU - Matkó, János AU - Drahos, László AU - Juhász, Gábor Dénes AU - Kékesi, Adrienna Katalin AU - Kardos, József TI - Local apoptotic-like mechanisms underlie complement-mediated synaptic pruning JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 115 PY - 2018 IS - 24 SP - 6303 EP - 6308 PG - 6 SN - 0027-8424 DO - 10.1073/pnas.1722613115 UR - https://m2.mtmt.hu/api/publication/3387341 ID - 3387341 N1 - ELTE NAP Neuroimmunology Research Group, Department of Biochemistry, Institute of Biology, ELTE Eötvös Loránd University, Budapest, H-1117, Hungary Laboratory of Proteomics, Institute of Biology, ELTE Eötvös Loránd University, Budapest, H-1117, Hungary Laboratory of Molecular Cell Biology, Institute of Enzymology, Hungarian Academy of Sciences, Budapest, H-1117, Hungary Department of Biochemistry, Institute of Biology, ELTE Eötvös Loránd University, Budapest, H-1117, Hungary MTA-TTK NAP B MS Neuroproteomics Group, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, H-1117, Hungary Department of Anatomy, Cell and Developmental Biology, Institute of Biology, ELTE Eötvös Loránd University, Budapest, H-1117, Hungary Laboratory of Human Brain Research, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, H-1083, Hungary Department of Immunology, Institute of Biology, ELTE Eötvös Loránd University, Budapest, H-1117, Hungary CRU Hungary Ltd., Göd, H-2131, Hungary Department of Physiology and Neurobiology, Institute of Biology, ELTE Eötvös Loránd University, Budapest, H-1117, Hungary Cited By :90 Export Date: 28 February 2023 CODEN: PNASA Correspondence Address: Kardos, J.; ELTE NAP Neuroimmunology Research Group, Hungary; email: kardos@elte.hu AB - C1q, a member of the immune complement cascade, is implicated in the selective pruning of synapses by microglial phagocytosis. C1q-mediated synapse elimination has been shown to occur during brain development, while increased activation and complement-dependent synapse loss is observed in neurodegenerative diseases. However, the molecular mechanisms underlying C1q-controlled synaptic pruning are mostly unknown. This study addresses distortions in the synaptic proteome leading to C1q-tagged synapses. Our data demonstrated the preferential localization of C1q to the presynapse. Proteomic investigation and pathway analysis of C1q-tagged synaptosomes revealed the presence of apoptotic-like processes in C1q-tagged synapses, which was confirmed experimentally with apoptosis markers. Moreover, the induction of synaptic apoptotic-like mechanisms in a model of sensory deprivation-induced synaptic depression led to elevated C1q levels. Our results unveiled that C1q label-based synaptic pruning is triggered by and directly linked to apoptotic-like processes in the synaptic compartment. LA - English DB - MTMT ER - TY - JOUR AU - Szebényi, Kornélia AU - Péntek, Adrienn AU - Erdei, Zsuzsa AU - Várady, György AU - Orbán, Tamás I. AU - Sarkadi, Balázs AU - Apáti, Ágota TI - Efficient Generation of Human Embryonic Stem Cell-Derived Cardiac Progenitors Based on Tissue-Specific Enhanced Green Fluorescence Protein Expression JF - TISSUE ENGINEERING PART C METHODS J2 - TISSUE ENG PART C METHODS VL - 21 PY - 2015 IS - 1 SP - 35 EP - 45 PG - 11 SN - 1937-3384 DO - 10.1089/ten.TEC.2013.0646 UR - https://m2.mtmt.hu/api/publication/2733708 ID - 2733708 AB - Cardiac progenitor cells (CPCs) are committed to the cardiac lineage but retain their proliferative capacity before becoming quiescent mature cardiomyocytes (CMs). In medical therapy and research, the use of human pluripotent stem cell-derived CPCs would have several advantages compared with mature CMs, as the progenitors show better engraftment into existing heart tissues, and provide unique potential for cardiovascular developmental as well as for pharmacological studies. Here, we demonstrate that the CAG promoter-driven EGFP reporter system enables the identification and isolation of embryonic stem cell-derived CPCs. Tracing of CPCs during differentiation confirmed up-regulation of surface markers, previously described to identify cardiac precursors and early CMs. Isolated CPCs express cardiac lineage-specific transcripts, still have proliferating capacity, and can be re-aggregated into embryoid body-like structures (CAG-EGFPhigh rEBs). Expression of troponin T and NKX2.5 mRNA is up-regulated in long-term cultured CAG-EGFPhigh rEBs, in which more than 90% of the cells become Troponin I positive mature CMs. Moreover, about one third of the CAG-EGFPhigh rEBs show spontaneous contractions. The method described here provides a powerful tool to generate expandable cultures of pure human CPCs that can be used for exploring early markers of the cardiac lineage, as well as for drug screening or tissue engineering applications. LA - English DB - MTMT ER -