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An improved transplantation strategy for mouse mesenchymal stem cells in an acute myocardial infarction model.

Jin J, Zhao Y, Tan X, Guo C, Yang Z, Miao D - PLoS ONE (2011)

Bottom Line: The 1(st) BM-MSCs maintained greater differentiation potentials towards cardiomocytes or vascular endothelial cells in vitro.This is indicated by higher expressions of cardiomyocyte and vascular endothelial cell mature markers in vitro.Furthermore, we identified that 24 proteins were down-regulated and 3 proteins were up-regulated in the 5(th) BM-MSCs in comparison to the 1(st) BM-MSCs, using mass spectrometry following two-dimensional electrophoresis.

View Article: PubMed Central - PubMed

Affiliation: The Research Center for Bone and Stem Cells, Department of Anatomy, Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu, The People's Republic of China.

ABSTRACT
To develop an effective therapeutic strategy for cardiac regeneration using bone marrow mesenchymal stem cells (BM-MSCs), the primary mouse BM-MSCs (1(st) BM-MSCs) and 5(th) passage BM-MSCs from β-galactosidase transgenic mice were respectively intramyocardially transplanted into the acute myocardial infarction (AMI) model of wild type mice. At the 6(th) week, animals/tissues from the 1(st) BM-MSCs group, the 5(th) passage BM-MSCs group, control group were examined. Our results revealed that, compared to the 5(th) passage BM-MSCs, the 1(st) BM-MSCs had better therapeutic effects in the mouse MI model. The 1(st) BM-MSCs maintained greater differentiation potentials towards cardiomocytes or vascular endothelial cells in vitro. This is indicated by higher expressions of cardiomyocyte and vascular endothelial cell mature markers in vitro. Furthermore, we identified that 24 proteins were down-regulated and 3 proteins were up-regulated in the 5(th) BM-MSCs in comparison to the 1(st) BM-MSCs, using mass spectrometry following two-dimensional electrophoresis. Our data suggest that transplantation of the 1(st) BM-MSCs may be an effective therapeutic strategy for cardiac tissue regeneration following AMI, and altered protein expression profiles between the 1(st) BM-MSCs and 5(th) passage BM-MSCs may account for the difference in their maintenance of stemness and their therapeutic effects following AMI.

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Protein expression profile and differentiation of the primary and 5th passaged BM-MSCs toward cardiocytes and vascular endothelial cells in vitro.(A) The representative images of silver stained 2-DE gel of the proteins from the primary pour-off cultures (upper panel, 1st BM-MSCs) and the 5th passaged cultures (low panel, 5th BM-MSCs). (B) Desmin, (C) Troponin I, (D) MHC-ß, (E) VEGF, (F) FLT-1 and (G) KDR mRNA levels were examined in BM-MSCs from the primary pour-off cultures (1st BM-MSCs) and the 5th passaged cultures (5th BM-MSCs) induced by 5′-azacytidine (B–D) and VEGF (E–G) by real-time RT-PCR. The mRNA levels were calculated as a ratio to the GAPDH mRNA level and expressed relative to levels of the 1st BM-MSCs. Each value was the means±S.E.M. of 5 determinations. **: P<0.01; ***: P<0.001 compared with the 1st BM-MSCs.
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pone-0021005-g003: Protein expression profile and differentiation of the primary and 5th passaged BM-MSCs toward cardiocytes and vascular endothelial cells in vitro.(A) The representative images of silver stained 2-DE gel of the proteins from the primary pour-off cultures (upper panel, 1st BM-MSCs) and the 5th passaged cultures (low panel, 5th BM-MSCs). (B) Desmin, (C) Troponin I, (D) MHC-ß, (E) VEGF, (F) FLT-1 and (G) KDR mRNA levels were examined in BM-MSCs from the primary pour-off cultures (1st BM-MSCs) and the 5th passaged cultures (5th BM-MSCs) induced by 5′-azacytidine (B–D) and VEGF (E–G) by real-time RT-PCR. The mRNA levels were calculated as a ratio to the GAPDH mRNA level and expressed relative to levels of the 1st BM-MSCs. Each value was the means±S.E.M. of 5 determinations. **: P<0.01; ***: P<0.001 compared with the 1st BM-MSCs.

Mentions: The comparison of protein expression profile was performed by 2-D electrophoresis (Figures 3A) and mass spectrometric analysis. Our results showed that 27 protein spots were identified to be differentially expressed by the 1st BM-MSCs for at least 5 times in comparison to the 5th BM-MSCs. Among those proteins, 24 proteins were down-regulated and 3 proteins were up-regulated in the 5th BM-MSCs. Some proteins among them have been known to be involved in regulating cellular functions of stem cells, which include adhesion, homing, engraftment, angiogenesis, anti-apoptosis, protecting cells from oxidative stress, and modulating inflammatory and immune responses (Table S1).


An improved transplantation strategy for mouse mesenchymal stem cells in an acute myocardial infarction model.

Jin J, Zhao Y, Tan X, Guo C, Yang Z, Miao D - PLoS ONE (2011)

Protein expression profile and differentiation of the primary and 5th passaged BM-MSCs toward cardiocytes and vascular endothelial cells in vitro.(A) The representative images of silver stained 2-DE gel of the proteins from the primary pour-off cultures (upper panel, 1st BM-MSCs) and the 5th passaged cultures (low panel, 5th BM-MSCs). (B) Desmin, (C) Troponin I, (D) MHC-ß, (E) VEGF, (F) FLT-1 and (G) KDR mRNA levels were examined in BM-MSCs from the primary pour-off cultures (1st BM-MSCs) and the 5th passaged cultures (5th BM-MSCs) induced by 5′-azacytidine (B–D) and VEGF (E–G) by real-time RT-PCR. The mRNA levels were calculated as a ratio to the GAPDH mRNA level and expressed relative to levels of the 1st BM-MSCs. Each value was the means±S.E.M. of 5 determinations. **: P<0.01; ***: P<0.001 compared with the 1st BM-MSCs.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3117862&req=5

pone-0021005-g003: Protein expression profile and differentiation of the primary and 5th passaged BM-MSCs toward cardiocytes and vascular endothelial cells in vitro.(A) The representative images of silver stained 2-DE gel of the proteins from the primary pour-off cultures (upper panel, 1st BM-MSCs) and the 5th passaged cultures (low panel, 5th BM-MSCs). (B) Desmin, (C) Troponin I, (D) MHC-ß, (E) VEGF, (F) FLT-1 and (G) KDR mRNA levels were examined in BM-MSCs from the primary pour-off cultures (1st BM-MSCs) and the 5th passaged cultures (5th BM-MSCs) induced by 5′-azacytidine (B–D) and VEGF (E–G) by real-time RT-PCR. The mRNA levels were calculated as a ratio to the GAPDH mRNA level and expressed relative to levels of the 1st BM-MSCs. Each value was the means±S.E.M. of 5 determinations. **: P<0.01; ***: P<0.001 compared with the 1st BM-MSCs.
Mentions: The comparison of protein expression profile was performed by 2-D electrophoresis (Figures 3A) and mass spectrometric analysis. Our results showed that 27 protein spots were identified to be differentially expressed by the 1st BM-MSCs for at least 5 times in comparison to the 5th BM-MSCs. Among those proteins, 24 proteins were down-regulated and 3 proteins were up-regulated in the 5th BM-MSCs. Some proteins among them have been known to be involved in regulating cellular functions of stem cells, which include adhesion, homing, engraftment, angiogenesis, anti-apoptosis, protecting cells from oxidative stress, and modulating inflammatory and immune responses (Table S1).

Bottom Line: The 1(st) BM-MSCs maintained greater differentiation potentials towards cardiomocytes or vascular endothelial cells in vitro.This is indicated by higher expressions of cardiomyocyte and vascular endothelial cell mature markers in vitro.Furthermore, we identified that 24 proteins were down-regulated and 3 proteins were up-regulated in the 5(th) BM-MSCs in comparison to the 1(st) BM-MSCs, using mass spectrometry following two-dimensional electrophoresis.

View Article: PubMed Central - PubMed

Affiliation: The Research Center for Bone and Stem Cells, Department of Anatomy, Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu, The People's Republic of China.

ABSTRACT
To develop an effective therapeutic strategy for cardiac regeneration using bone marrow mesenchymal stem cells (BM-MSCs), the primary mouse BM-MSCs (1(st) BM-MSCs) and 5(th) passage BM-MSCs from β-galactosidase transgenic mice were respectively intramyocardially transplanted into the acute myocardial infarction (AMI) model of wild type mice. At the 6(th) week, animals/tissues from the 1(st) BM-MSCs group, the 5(th) passage BM-MSCs group, control group were examined. Our results revealed that, compared to the 5(th) passage BM-MSCs, the 1(st) BM-MSCs had better therapeutic effects in the mouse MI model. The 1(st) BM-MSCs maintained greater differentiation potentials towards cardiomocytes or vascular endothelial cells in vitro. This is indicated by higher expressions of cardiomyocyte and vascular endothelial cell mature markers in vitro. Furthermore, we identified that 24 proteins were down-regulated and 3 proteins were up-regulated in the 5(th) BM-MSCs in comparison to the 1(st) BM-MSCs, using mass spectrometry following two-dimensional electrophoresis. Our data suggest that transplantation of the 1(st) BM-MSCs may be an effective therapeutic strategy for cardiac tissue regeneration following AMI, and altered protein expression profiles between the 1(st) BM-MSCs and 5(th) passage BM-MSCs may account for the difference in their maintenance of stemness and their therapeutic effects following AMI.

Show MeSH
Related in: MedlinePlus