ABSTRACT
Objective To investigate the efficiency of target gene transfection of the heart and liver after tail vein or intramyocardial injection of adenovirus vector (GFP-Ad).Methods GFP-AD was constructed at first.A total of 20 male 8-week old C57BL/6 mice were randomly and equally divided into tail vein injection of GFP-AD group and intramyocardial injection of GFP-AD group.The mRNA levels of GFP in the heart and liver tissues were detected by Q-PCR at different time points.Fluorescence microscopy was performed to visualize the expression of GFP fluorescence.Results Compared with the tail vein injection group, the GFP mRNA level in mouse heart tissue was apparently higher in the intramyocardial injection group.In both groups, the GFP mRNA levels in liver tissue were significantly increased compared with that in the heart tissue.In the tail vein injection group, the GFP mRNA level in liver tissue reached a peak on day 7;but in the intramyocardial injection group, the mRNA level of GFP in liver tissue reached apeak on day 3.We also observed the same trend of GFP fluorescence expression in the tail vein injection group compared with that in the intramyocardial injection group.Conclusions Intramyocardial injection of adenovirus vector is suitable to achieve a higher transfection efficiency in mouse heart tissue compared with the tail vein injection method.Although both injection methods are suitable for transfection of mouse liver, the tail vein injection method is preferential for it is simple and less invasive.
ABSTRACT
O uso de células-tronco representa uma alternativa para o tratamento das doenças que acometem o coração, devido à capacidade que essas células indiferenciadas têm de preservar sua própria população e de se diferenciar em células dos diversos tecidos, incluindo o cardíaco. Nesse trabalho comparamos as características de células-tronco isoladas a partir do tecido cardíaco e da medula óssea de camundongos transgênicos para a proteína fluorescente verde (GFP). As células-tronco cardíacas e da medula óssea apresentaram característica morfológica fibroblastóide e imunofenotípica de células-tronco mesenquimais, com alta expressão dos marcadores CD44, CD90, CD73, Sca-1 e baixa expressão dos marcadores de células hematopoiéticas. A análise citogenética revelou um cariótipo poliplóide a partir da terceira passagem das células-tronco isoladas do coração e da medula-óssea. A capacidade de diferenciação em vários tipos celulares, tais como adipócitos, osteócitos e condrócitos, também foi avaliada nas células-tronco de ambas as fontes. Tanto as células-tronco isoladas do coração como da medula óssea foram capazes de se diferenciar nessas três linhagens. Quando estimuladas com 5azacitidina para testar o potencial cardiomiogênico das células isoladas do coração e da medula óssea, apenas as células-tronco cardíacas passaram a expressar alguns marcadores de cardiomiócitos, tais como troponina T cardíaca e GATA-4...
Stem cells are undifferentiated cells with the ability of self-renewal and differentiation into different cell types, with the potential to treat heart diseases. In the present study we compared the characteristics of stem cells isolated from the heart to bone marrow stem cells, both obtained from EGFP transgeneic mice. Cardiac and bone marrow stem cells presented fibroblastic morphology and an immunophenotype compatible with mesenchymal stem cells high expression of CD44, CD90, CD73, Sca-1 and low expression of of hematopoietic lineage markers. Cytogenetic analysis demonstrated polyploid karyotypes after the third passage of the stem cells isolated from heart and bone marrow. Both bone marrow and heart stem cells were able to differentiate into adipocytes, osteocytes and chondrocytes. In order to test the potential of differentiation into cardiomyocytes, cells were stimulated with 5azacytidine and only cardiac stem cells expressed heart-specific markers: cardiac T troponin and GATA-4...