Kutschka, Ingo

[["dc.bibliographiccitation.artnumber","e0192652"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","PLOS ONE"],["dc.bibliographiccitation.volume","13"],["dc.contributor.author","Dahlmann, Julia"],["dc.contributor.author","Awad, George"],["dc.contributor.author","Dolny, Carsten"],["dc.contributor.author","Weinert, Sönke"],["dc.contributor.author","Richter, Karin"],["dc.contributor.author","Fischer, Klaus-Dieter"],["dc.contributor.author","Munsch, Thomas"],["dc.contributor.author","Leßmann, Volkmar"],["dc.contributor.author","Volleth, Marianne"],["dc.contributor.author","Zenker, Martin"],["dc.contributor.author","Chen, Yaoyao"],["dc.contributor.author","Merkl, Claudia"],["dc.contributor.author","Schnieke, Angelika"],["dc.contributor.author","Baraki, Hassina"],["dc.contributor.author","Kutschka, Ingo"],["dc.contributor.author","Kensah, George"],["dc.date.accessioned","2019-07-09T11:45:08Z"],["dc.date.available","2019-07-09T11:45:08Z"],["dc.date.issued","2018"],["dc.description.abstract","The possibility to generate cardiomyocytes from pluripotent stem cells in vitro has enormous significance for basic research, disease modeling, drug development and heart repair. The concept of heart muscle reconstruction has been studied and optimized in the rat model using rat primary cardiovascular cells or xenogeneic pluripotent stem cell derived-cardiomyocytes for years. However, the lack of rat pluripotent stem cells (rPSCs) and their cardiovascular derivatives prevented the establishment of an authentic clinically relevant syngeneic or allogeneic rat heart regeneration model. In this study, we comparatively explored the potential of recently available rat embryonic stem cells (rESCs) and induced pluripotent stem cells (riPSCs) as a source for cardiomyocytes (CMs). We developed feeder cell-free culture conditions facilitating the expansion of undifferentiated rPSCs and initiated cardiac differentiation by embryoid body (EB)-formation in agarose microwell arrays, which substituted the robust but labor-intensive hanging drop (HD) method. Ascorbic acid was identified as an efficient enhancer of cardiac differentiation in both rPSC types by significantly increasing the number of beating EBs (3.6 ± 1.6-fold for rESCs and 17.6 ± 3.2-fold for riPSCs). These optimizations resulted in a differentiation efficiency of up to 20% cTnTpos rPSC-derived CMs. CMs showed spontaneous contractions, expressed cardiac markers and had typical morphological features. Electrophysiology of riPSC-CMs revealed different cardiac subtypes and physiological responses to cardio-active drugs. In conclusion, we describe rPSCs as a robust source of CMs, which is a prerequisite for detailed preclinical studies of myocardial reconstruction in a physiologically and immunologically relevant small animal model."],["dc.identifier.doi","10.1371/journal.pone.0192652"],["dc.identifier.pmid","29513687"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/15042"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/59166"],["dc.language.iso","en"],["dc.notes.intern","Merged from goescholar"],["dc.relation","info:eu-repo/grantAgreement/EC/FP7/241504/EU//EURATRANS"],["dc.relation.issn","1932-6203"],["dc.rights","CC BY 4.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.subject.ddc","610"],["dc.title","Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture."],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","96"],["dc.bibliographiccitation.issue","1"],["dc.bibliographiccitation.journal","Cell Communication and Signaling"],["dc.bibliographiccitation.volume","16"],["dc.contributor.author","Haghighi, Fereshteh"],["dc.contributor.author","Dahlmann, Julia"],["dc.contributor.author","Nakhaei-Rad, Saeideh"],["dc.contributor.author","Lang, Alexander"],["dc.contributor.author","Kutschka, Ingo"],["dc.contributor.author","Zenker, Martin"],["dc.contributor.author","Kensah, George"],["dc.contributor.author","Piekorz, Roland P"],["dc.contributor.author","Ahmadian, Mohammad R"],["dc.date.accessioned","2019-07-09T11:49:36Z"],["dc.date.available","2019-07-09T11:49:36Z"],["dc.date.issued","2018"],["dc.description.abstract","Abstract Background Human pluripotent stem cells (PSCs) open new windows for basic research and regenerative medicine due to their remarkable properties, i.e. their ability to self-renew indefinitely and being pluripotent. There are different, conflicting data related to the role of basic fibroblast growth factor (bFGF) in intracellular signal transduction and the regulation of pluripotency of PSCs. Here, we investigated the effect of bFGF and its downstream pathways in pluripotent vs. differentiated human induced (hi) PSCs. Methods bFGF downstream signaling pathways were investigated in long-term culture of hiPSCs from pluripotent to differentiated state (withdrawing bFGF) using immunoblotting, immunocytochemistry and qPCR. Subcellular distribution of signaling components were investigated by simple fractionation and immunoblotting upon bFGF stimulation. Finally, RAS activity and RAS isoforms were studied using RAS assays both after short- and long-term culture in response to bFGF stimulation. Results Our results revealed that hiPSCs were differentiated into the ectoderm lineage upon withdrawing bFGF as an essential pluripotency mediator. Pluripotency markers OCT4, SOX2 and NANOG were downregulated, following a drastic decrease in MAPK pathway activity levels. Notably, a remarkable increase in phosphorylation levels of p38 and JAK/STAT3 was observed in differentiated hiPSCs, while the PI3K/AKT and JNK pathways remained active during differentiation. Our data further indicate that among the RAS paralogs, NRAS predominantly activates the MAPK pathway in hiPSCs. Conclusion Collectively, the MAPK pathway appears to be the prime signaling pathway downstream of bFGF for maintaining pluripotency in hiPSCs and among the MAPK pathways, the activity of NRAS-RAF-MEK-ERK is decreased during differentiation, whereas p38 is activated and JNK remains constant."],["dc.identifier.doi","10.1186/s12964-018-0307-1"],["dc.identifier.purl","https://resolver.sub.uni-goettingen.de/purl?gs-1/15725"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/59591"],["dc.language.iso","en"],["dc.notes.intern","Merged from goescholar"],["dc.publisher","BioMed Central"],["dc.rights","CC BY 4.0"],["dc.rights.uri","https://creativecommons.org/licenses/by/4.0"],["dc.title","bFGF-mediated pluripotency maintenance in human induced pluripotent stem cells is associated with NRAS-MAPK signaling"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.version","published_version"],["dspace.entity.type","Publication"]]