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Eur J Cardiothorac Surg 2005;28:685-691
© 2005 Elsevier Science NL

Fusion of bone marrow-derived stem cells with cardiomyocytes in a heterologous in vitro model

^a Department of Cardiac Surgery, Heart Center, University of Leipzig, Struempellstr. 39, 04289 Leipzig, Germany
^b Department of Paediatric Cardiology, Heart Center, University of Leipzig, Germany

Received 26 November 2004; received in revised form 17 June 2005; accepted 27 June 2005.

^_* Corresponding author. Tel.: +49 341 8651420; fax: +49 341 8651452. (Email: jgmed93{at}hotmail.com).

OBJECTIVE: Recent studies have demonstrated that transplanted bone marrow-derived stem cells (BMCs) possess a broad differentiation potential and are able to form new cardiomyocytes. However, the identity of BMCs as true cardiomyocytes is still ambiguous. Therefore, we investigated the fate of transplanted fluorescence labeled BMCs and cardiomyocytes in co-culture. Methods: For cell tracking we used two different fluorescent probes, Vybrant/DiO and Vybrant/DiI. BMCs were taken from human sternal marrow, purified using a Ficoll-gradient-centrifugation, treated with 5-azacytidine and stained with Vybrant/DiO. Furthermore, isolated spontaneous beating cardiomyocytes of neonatal rats (CM) were labeled with Vybrant/DiI. Thereafter, the BMCs were transplanted into CM-cultures and investigated on day 1, 4, 7, 14 and 28 using two-color fluorescence phenotyping by laser-scanning-cytometry (LSC). Two-color positive cells were harvested by patch–clamp technique and ß-MHC mRNA expression was analyzed by single-cell PCR. Results: Two different morphological phenotypes were observed by LSC. First, isolated DiO labeled BMCs without contact or with direct cell contact to DiI labeled CMs. Second, some BMCs and CMs were double positive for DiO/DiI spontaneously forming hybrids. This population increased by 18% from day 1 to 4 and decreased only slightly until day 28. Additionally, few two-color positive cell formations expressed both human and rat specific ß-MHC mRNA as well as only human ß-MHC mRNA indicating that cell-fusion and transdifferentiation has occurred. Conclusion: These observations provide in vitro evidence for spontaneous cell fusion and transdifferentiation of BMCs in co-culture, raising the possibility that the observed phenomenons may contribute to development or maintenance of these cell types.

Key Words: Bone marrow stem cells • Cell fusion • Transdifferentiation • Heart failure • Myocardial regeneration

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