Coculture with hematopoietic stem cells protects cardiomyocytes against apoptosis via paracrine activation of AKT

Journal of Translational Medicine

Norbert Frey²  Hugo A Katus¹  Jutta Krebs¹  Volker Eckstein³  Rainer Will¹  Constantin Kühl²  Matthias Lutz²  Mark Rosenberg² 

[1] Department of Internal Medicine III, University of Heidelberg, Heidelberg, Germany;Department of Internal Medicine III (Cardiology and Angiology), University Medical Center Schleswig-Holstein, Campus Kiel, Schittenhelmstr.12, D-24105, Kiel, Germany;Department of Internal Medicine V, University of Heidelberg, Heidelberg, Germany

关键词: Paracrine;    Apoptosis;    Cardiomyocytes;    Stem cells;   

Others  :  1205966
DOI  :  10.1186/1479-5876-10-115

 received in 2011-09-22, accepted in 2012-06-06,  发布年份 2012

【 摘 要 】

Background

Previous experimental studies concluded that stem cells (SC) may exert their beneficial effects on the ischemic heart by paracrine activation of antiapoptotic pathways. In order to identify potential cardioprotective mediators, we performed a systematic analysis of the differential gene expression of hematopoietic SC after coculture with cardiomyocytes (CM).

Methods

After 48 h of coculture with neonatal rat ventricular CM (NRVCM), two consecutive cell sorting steps generated a highly purified population of conditioned murine hematopoietic SC (>99%). Next, a genome-wide microarray analysis of cocultured vs. monocultured hematopoietic SC derived from three independent experiments was performed. The analysis of differentially expressed genes was focused on products that are secretable and/or membrane-bound and potentially involved in antiapoptotic signalling.

Results

We found CCL-12, Macrophage Inhibitory Factor, Fibronectin and connexin 40 significantly upregulated in our coculture model. An ELISA of cell culture supernatants was performed to confirm secretion of candidate genes and showed that coculture supernatants revealed markedly higher CCL-12 concentrations. Moreover, we stimulated NRVCM with concentrated coculture supernatants which resulted in a significant reduction of apoptosis compared to monoculture-derived supernatant. Mechanistically, NRVCMs stimulated with coculture supernatants showed a higher level of AKT-phosphorylation, consistent with enhanced antiapoptotic signaling.

Conclusion

In summary, our results show that the interaction between hematopoietic SC and NRVCM led to a modified gene expression and induction of antiapoptotic pathways. These findings may thus at least in part explain the cardioprotective effects of hematopoietic SC.

【 授权许可】

   
2012 Rosenberg et al.; licensee BioMed Central Ltd.

【 图 表 】

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