In-vitro-Charakterisierung und kardiale Differenzierung von induziert pluripotenten Stammzellen der Maus

Lentzen, Max-Philipp

dc.contributor.advisor	Becker, Alexander Dr.
dc.contributor.author	Lentzen, Max-Philipp
dc.date.accessioned	2016-04-04T08:06:57Z
dc.date.available	2016-04-13T22:50:05Z
dc.date.issued	2016-04-04
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-0028-8716-4
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-5594
dc.language.iso	deu	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	610	de
dc.title	In-vitro-Charakterisierung und kardiale Differenzierung von induziert pluripotenten Stammzellen der Maus	de
dc.type	doctoralThesis	de
dc.title.translated	In vitro characterisation and cardiac differentiation of murine induced pluripotent stem cells	de
dc.contributor.referee	Guan-Schmidt, Kaomei Prof. Dr.
dc.date.examination	2016-04-06
dc.description.abstracteng	The following paper is part of many recent research projects that try to find and establish a sufficient cell replacement therapy for structural myocardial deseases. Induced pluripotent stem cells which are comparable to embryonic stem cells and differentiate into cardiomyocytes seem to be an option. The task was to compare miPS with mES by characterisation and differentiation into cardiomyocytes and also characterise them. This was followed by antibiotic selection and purification via immunofluorescence. The in-vitro-results showed a successfull reprogramming with the transcription factors OCT4, SOX2, KLF4 und c-MYC in a single lentiviral stem cell cassette and also that the investigated miPS showed similar characteristics as the investigated ES. The proof of pluripotency showed no differences between the embryonic and the induced pluripotent stem cells neither on mRNA nor on protein level. The morphology of the cells during the differentiation of the cells of the three germ layers showed only minimal differences. There was also a physiological caryotype shown even after long term cultivation. By using a standardized EB based differentiation protocol (hanging drop-method) for ES the induced pluripotent and embryonic stem cells have been differentiated into cardiomyocytes over 22 days. The source of the investigated cells were double transgene mice (Mhc-Neo/Mhc-eGfp). With them it was possible to achieve a mhc dependent expression of eGfp and a neomycine resistance of the cardiomyocytes which made a selection and visualisation possible. Analysis of the molecular, structural and functional aspects of the iPS derived cardiomyocytes showed typical characteristics of ES derived cardiomyocytes. Investigations by rt-PCR showed the expression of typical mesodermal marker genes as Gata4, Nkx2.5, α-Mhc, Mlc2v and Anf. Also on protein level the expression of cardiac markers could be proofed. Thus the differentiation of induced pluripotent stem cells into functional cardiomyocytes could be demonstrated. By Use of iPS derived and not ES derived cardiomyocytes problems as immunological reactions or embryo consuming research as an ethical issue could be solved. Based on the results of this research project following investigations have to show if the generated cardiomyocytes are suitable for transplantation, if they survive and if they functional integrate in vivo. For a therapeutical application the achieved results also have to be reproduced in humans.	de
dc.contributor.coReferee	Dressel, Ralf Prof. Dr.
dc.subject.eng	cardiomyocytes	de
dc.subject.eng	induced pluripotent stem cells	de
dc.subject.eng	embryonic stem cells	de
dc.subject.eng	transcription factors	de
dc.subject.eng	lentiviral stem cell cassette	de
dc.subject.eng	pluripotency	de
dc.subject.eng	hanging drop	de
dc.subject.eng	double transgene mice	de
dc.subject.eng	neomycine	de
dc.subject.eng	eGfp	de
dc.identifier.urn	urn:nbn:de:gbv:7-11858/00-1735-0000-0028-8716-4-5
dc.affiliation.institute	Medizinische Fakultät	de
dc.subject.gokfull	Medizin (PPN619874732)	de
dc.subject.gokfull	Innere Medizin - Allgemein- und Gesamtdarstellungen (PPN619875747)	de
dc.description.embargoed	2016-04-13
dc.identifier.ppn	856161470

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