Universität zu Köln

Bader, Andreas Matthaeus, Brodarac, Andreja, Klose, Kristin, Bieback, Karen, Choi, Yeong-Hoon, Kurtz, Andreas ORCID: 0000-0003-3301-6546 and Stamm, Christof (2014). Mechanisms of paracrine cardioprotection by cord blood mesenchymal stromal cells. Eur. J. Cardio-Thorac. Surg., 45 (6). S. 983 - 993. CARY: OXFORD UNIV PRESS INC. ISSN 1873-734X

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Abstract

OBJECTIVES: Among the mechanisms by which somatic stem cells may improve left ventricular function in ischaemic heart disease are pro-survival stimuli mediated by secreted factors. This phenomenon is frequently referred to, but remains poorly understood. We therefore investigated the non-regenerative cardioprotective effects of cord blood mesenchymal stromal cells (CBMSCs) in vitro and sought to identify relevant intracellular signalling pathways. METHODS: Conditioned medium from CBMSCs and fibroblasts was prepared, and secreted factors were analysed by Luminex (R) immuno-bead assay. Murine cardiomyocyte-derived HL-1 cells were subjected to simulated ischaemia by glucose and serum deprivation and hypoxia in CBMSC-conditioned or cell-free control medium or in medium conditioned by foreskin fibroblasts. The proportions of vital, apoptotic and necrotic cells (poly-caspase activity, annexin V and ethidium homodimer-III staining) were quantified using a high-content imaging system. Metabolic activity and proliferation rate were determined via 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium and 5-bromo-2-deoxyuridine assays. Phosphorylation of Akt, extracellular-signal-regulated kinase (ERK) 1/2, signal transducer and activator of transcription 3 (STAT3) and glycogen synthase kinase 3 beta was determined by western blot, and experiments were repeated in the presence of specific small-molecule inhibitors (Wortmannin, UO126 and Stattic). RESULTS: CBMSC medium reduced the proportion of dead HL-1 cardiomyocytes from 39 +/- 3 to 28 +/- 1% (P < 0.05) and the rate of late apoptotic cells to 68 +/- 2% of that in control medium (P < 0.001). Metabolic activity was increased by 12 +/- 1% compared with control (P < 0.05), while in fibroblast medium it was not (5 +/- 2%, P = 1). This was associated with increased phosphorylation of Akt (2-fold, P < 0.05), ERK1/2 (3-fold, P < 0.01) and STAT3 (12-fold, P < 0.001). Combined blocking of the phosphatidylinositol-4,5-bisphosphate 3-kinase/Akt and mitogen-activated protein kinase/ERK signalling abolished the protective CBMSC effect, while blocking the pathways individually had no effect. Inhibition of STAT3 phosphorylation drastically lowered HL-1 cell viability in control medium, but not in medium conditioned by CBMSCs. CONCLUSIONS: The factors released by CBMSCs protect cardiomyocyte-like HL-1 cells from simulated ischaemia more than those released from fibroblasts. While CBMSC-triggered Akt and ERK1/2 activation provides protection in a compensatory manner, STAT3 is crucial for cardiomyocyte survival in ischaemia, but is not a key mediator of cytoprotective stem cell actions.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Bader, Andreas Matthaeus UNSPECIFIED UNSPECIFIED UNSPECIFIED Brodarac, Andreja UNSPECIFIED UNSPECIFIED UNSPECIFIED Klose, Kristin UNSPECIFIED UNSPECIFIED UNSPECIFIED Bieback, Karen UNSPECIFIED UNSPECIFIED UNSPECIFIED Choi, Yeong-Hoon UNSPECIFIED UNSPECIFIED UNSPECIFIED Kurtz, Andreas UNSPECIFIED orcid.org/0000-0003-3301-6546 UNSPECIFIED Stamm, Christof UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-436929
DOI:	10.1093/ejcts/ezt576
Journal or Publication Title:	Eur. J. Cardio-Thorac. Surg.
Volume:	45
Number:	6
Page Range:	S. 983 - 993
Date:	2014
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 BONE-MARROW-CELLS; STEM-CELLS; MYOCARDIAL-INFARCTION; IN-VITRO; SURVIVAL; REPERFUSION; ACTIVATION; THERAPY; STAT3 Multiple languages Cardiac & Cardiovascular Systems; Respiratory System; Surgery Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/43692