Universität zu Köln

Oberwallner, Barbara, Brodarac, Andreja, Anic, Petra, Saric, Tomo ORCID: 0000-0001-8344-1095, Wassilew, Katharina ORCID: 0000-0002-8683-465X, Neef, Klaus, Choi, Yeong-Hoon and Stamm, Christof (2015). Human cardiac extracellular matrix supports myocardial lineage commitment of pluripotent stem cells. Eur. J. Cardio-Thorac. Surg., 47 (3). S. 416 - 426. CARY: OXFORD UNIV PRESS INC. ISSN 1873-734X

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Abstract

OBJECTIVES: Cross-talk between organ-specific extracellular matrix (ECM) and stem cells is often assumed but has not been directly demonstrated. We developed a protocol for the preparation of human cardiac ECM (cECM) and studied whether cECM has effects on pluripotent stem cell differentiation that may be useful for future cardiac regeneration strategies in patients with end-stage heart failure. METHODS: Of note, 0.3 mm-thick cECM slices were prepared from samples of myocardium from patients with end-stage non-ischaemic dilated cardiomyopathy, using a three-step protocol involving hypotonic lysis buffer, sodium dodecyl sulphate (SDS) and foetal bovine serum (FBS). Murine embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs) and mesenchymal stromal cells (MSCs) were seeded and grown in standard culture, on cECM or on non-specific ECM preparations (Matrigel (R) or Geltrex (R)). Cell attachment, apoptosis induction (Caspase 3/7 activity) and metabolic activity (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium conversion) were followed. Transcriptional activation of genes involved in pluripotency; early and late myocardial development; and endothelial, ectodermal or endodermal commitment were monitored by quantitative real-time polymerase chain reaction (rtPCR). Protein expression of selected markers was confirmed by immunohistology. RESULTS: cECM supported the proliferation of ESCs and iPSCs, and Caspase 3/7 activity was significantly lower compared with standard culture. Cardiac lineage commitment was favoured when ESCs or iPSCs were grown on cECM, as evidenced by the significantly increased mRNA expression of cardiac alpha myosin heavy polypeptide 6 (Myh6), cardiac troponin T2 (Tnnt2) and NK2 homeobox 5 (Nkx2.5) as well as positive immunohistology for cardiac troponin T and heavy-chain cardiac myosin protein. In contrast, Matrigel or Geltrex did not induce cardiac-specific markers. MSCs showed no evidence of cardiomyocyte differentiation. CONCLUSIONS: Human cardiac ECM seems to direct differentiation of pluripotent stem cells towards a cardiomyocyte phenotype. This phenomenon supports the use of cardiac ECM preparations for guided stem cell differentiation and myocardial repair, and may ultimately increase the therapeutic efficacy of cell therapy in heart failure patients.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Oberwallner, Barbara UNSPECIFIED UNSPECIFIED UNSPECIFIED Brodarac, Andreja UNSPECIFIED UNSPECIFIED UNSPECIFIED Anic, Petra UNSPECIFIED UNSPECIFIED UNSPECIFIED Saric, Tomo UNSPECIFIED orcid.org/0000-0001-8344-1095 UNSPECIFIED Wassilew, Katharina UNSPECIFIED orcid.org/0000-0002-8683-465X UNSPECIFIED Neef, Klaus UNSPECIFIED UNSPECIFIED UNSPECIFIED Choi, Yeong-Hoon UNSPECIFIED UNSPECIFIED UNSPECIFIED Stamm, Christof UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-410304
DOI:	10.1093/ejcts/ezu163
Journal or Publication Title:	Eur. J. Cardio-Thorac. Surg.
Volume:	47
Number:	3
Page Range:	S. 416 - 426
Date:	2015
Publisher:	OXFORD UNIV PRESS INC
Place of Publication:	CARY
ISSN:	1873-734X
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language DIFFERENTIATION; CARDIOMYOCYTES; EXPRESSION; PHENOTYPE; SCAFFOLDS; PLATFORM; HEART; MODEL Multiple languages Cardiac & Cardiovascular Systems; Respiratory System; Surgery Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/41030