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Zimmermann, Martina E. ; Fuerstenau-Sharp, Maya ; Stark, Klaus ; Jentsch, Nico ; Klingenstein, Melanie ; Drzymalski, Marzena ; Wagner, Stefan ; Maier, Lars S. ; Baessler, Andrea ; Fischer, Marcus ; Hengstenberg, Christian ; Hehr, Ute

Generation of Highly Purified Human Cardiomyocytes from Peripheral Blood Mononuclear Cell-Derived Induced Pluripotent Stem Cells

Zimmermann, Martina E., Fuerstenau-Sharp, Maya, Stark, Klaus, Jentsch, Nico, Klingenstein, Melanie, Drzymalski, Marzena, Wagner, Stefan, Maier, Lars S., Baessler, Andrea, Fischer, Marcus, Hengstenberg, Christian und Hehr, Ute (2015) Generation of Highly Purified Human Cardiomyocytes from Peripheral Blood Mononuclear Cell-Derived Induced Pluripotent Stem Cells. PLoS ONE 10 (5), S. 1-21.

Veröffentlichungsdatum dieses Volltextes: 18 Jun 2015 15:43
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.31972


Zusammenfassung

Induced pluripotent stem (iPS) cells have an enormous potential for physiological studies. A novel protocol was developed combining the derivation of iPS from peripheral blood with an optimized directed differentiation to cardiomyocytes and a subsequent metabolic selection. The human iPS cells were retrovirally dedifferentiated from activated T cells. The subsequent optimized directed ...

Induced pluripotent stem (iPS) cells have an enormous potential for physiological studies. A novel protocol was developed combining the derivation of iPS from peripheral blood with an optimized directed differentiation to cardiomyocytes and a subsequent metabolic selection. The human iPS cells were retrovirally dedifferentiated from activated T cells. The subsequent optimized directed differentiation protocol yielded 30-45% cardiomyocytes at day 16 of differentiation. The derived cardiomyocytes expressed appropriate structural markers like cardiac troponin T, alpha-actinin and myosin light chain 2 (MLC2V). In a subsequent metabolic selection with lactate, the cardiomyocytes content could be increased to more than 90%. Loss of cardiomyocytes during metabolic selection were less than 50%, whereas alternative surface antibody-based selection procedures resulted in loss of up to 80% of cardiomyocytes. Electrophysiological characterization confirmed the typical cardiac features and the presence of ventricular, atrial and nodal-like action potentials within the derived cardio-myocyte population. Our combined and optimized protocol is highly robust and applicable for scalable cardiac differentiation. It provides a simple and cost-efficient method without expensive equipment for generating large numbers of highly purified, functional cardiomyocytes. It will further enhance the applicability of iPS cell-derived cardiomyocytes for disease modeling, drug discovery, and regenerative medicine.



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Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftPLoS ONE
Verlag:PUBLIC LIBRARY SCIENCE
Ort der Veröffentlichung:SAN FRANCISCO
Band:10
Nummer des Zeitschriftenheftes oder des Kapitels:5
Seitenbereich:S. 1-21
Datum13 Mai 2015
InstitutionenMedizin > Lehrstuhl für Humangenetik
Medizin > Lehrstuhl für Innere Medizin II
Medizin > Institut für Epidemiologie und Präventivmedizin
Identifikationsnummer
WertTyp
10.1371/journal.pone.0126596DOI
Article ID: e0126596Andere
Stichwörter / KeywordsLONG-QT SYNDROME; ACTION-POTENTIALS; PROGENITOR CELLS; T-CELLS; MOUSE; CONTRACTION; DIFFERENTIATION; PURIFICATION; CURRENTS; MYOCYTES;
Dewey-Dezimal-Klassifikation600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin
StatusVeröffentlicht
BegutachtetJa, diese Version wurde begutachtet
An der Universität Regensburg entstandenJa
URN der UB Regensburgurn:nbn:de:bvb:355-epub-319725
Dokumenten-ID31972

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