Background: Mesenchymal stem cells (MSC) repair infarcted hearts mainly through the release of cytoprotective factors. We compared fetal MSC (F-MSC) with adult MSC from young and elderly patients to establish if donor age influences their cardioprotective paracrine properties. Methods: F-MSC were isolated from human placenta while adult MSC were isolated from bone marrow samples of young (yBM-MSC; age < 65 years) or old (oBM-MSC; age > 65 ) donors. Rat neonatal cardiomyocytes (H9c2) were exposed to hypoxia/reoxygenation (6/18 hours) in the presence of control medium (CTRL-M) or conditioned medium from F-MSC (F-CM), yBM-MSC (Y-CM) or oBM-MSC (O-CM). H9c2 viability was evaluated by MTS assay. Apoptosis was measured by TUNEL staining and by Caspase 3 activation (colorimetric assay and Western Blot). We used RT-PCR to verify the expression of known cytoprotective factors. Results: The hypoxia/reoxygenation protocol reduced H9c2 viability by 55% compared with basal condition (p<0.001). F-CM and Y-CM increased cell viability by 45% and 33% (p<0.017), respectively; O-CM had no significant effect on H9c2 viability (p=n.s. vs CTRL-M). Compared with CTRL-M and O-CM, F-CM significantly reduced the number of TUNEL positive nuclei by 91% (p<0.001) and 89% (p<0.001), respectively. The Y-CM also reduced H9c2 apoptotic nuclei (- 67,5% vs CTRL-M, p<0.01; - 64% vs O-CM, p<0.01). In contrast, O-CM did not prevent H9c2 apoptotic death (-11% vs CTRL-M, p=n.s.). Both colorimetric assay and Western Blot analysis showed that Caspase-3 activation was prevented by F-CM and Y-CM but not by O-CM. F-MSC expressed PDGF-β, BMP2, EPO, FGF2 and VEGF at significantly higher level compared with oBM-MSC (p<0.05); VEGF, FGF2 and HGF transcripts were significantly higher in yBM-MSC than in oBM-MSC (p<0.05). Conclusions: MSC can mediate cardiomyocyte protection through the release of soluble anti-apoptotic factors. However, we documented that donor age negatively influences the paracrine cytoprotective properties of adult MSC.
The cardioprotective paracrine effects exerted by human mesenchymal stem cells are negatively influenced by donor age
DANIELI, PATRIZIA;CERVIO, ELISABETTA;CIUFFREDA, MARIA CHIARA;PISANO, FEDERICA;ROCCIO, MARIANNA;GNECCHI, MASSIMILIANO
2010-01-01
Abstract
Background: Mesenchymal stem cells (MSC) repair infarcted hearts mainly through the release of cytoprotective factors. We compared fetal MSC (F-MSC) with adult MSC from young and elderly patients to establish if donor age influences their cardioprotective paracrine properties. Methods: F-MSC were isolated from human placenta while adult MSC were isolated from bone marrow samples of young (yBM-MSC; age < 65 years) or old (oBM-MSC; age > 65 ) donors. Rat neonatal cardiomyocytes (H9c2) were exposed to hypoxia/reoxygenation (6/18 hours) in the presence of control medium (CTRL-M) or conditioned medium from F-MSC (F-CM), yBM-MSC (Y-CM) or oBM-MSC (O-CM). H9c2 viability was evaluated by MTS assay. Apoptosis was measured by TUNEL staining and by Caspase 3 activation (colorimetric assay and Western Blot). We used RT-PCR to verify the expression of known cytoprotective factors. Results: The hypoxia/reoxygenation protocol reduced H9c2 viability by 55% compared with basal condition (p<0.001). F-CM and Y-CM increased cell viability by 45% and 33% (p<0.017), respectively; O-CM had no significant effect on H9c2 viability (p=n.s. vs CTRL-M). Compared with CTRL-M and O-CM, F-CM significantly reduced the number of TUNEL positive nuclei by 91% (p<0.001) and 89% (p<0.001), respectively. The Y-CM also reduced H9c2 apoptotic nuclei (- 67,5% vs CTRL-M, p<0.01; - 64% vs O-CM, p<0.01). In contrast, O-CM did not prevent H9c2 apoptotic death (-11% vs CTRL-M, p=n.s.). Both colorimetric assay and Western Blot analysis showed that Caspase-3 activation was prevented by F-CM and Y-CM but not by O-CM. F-MSC expressed PDGF-β, BMP2, EPO, FGF2 and VEGF at significantly higher level compared with oBM-MSC (p<0.05); VEGF, FGF2 and HGF transcripts were significantly higher in yBM-MSC than in oBM-MSC (p<0.05). Conclusions: MSC can mediate cardiomyocyte protection through the release of soluble anti-apoptotic factors. However, we documented that donor age negatively influences the paracrine cytoprotective properties of adult MSC.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
