Hepatic stellate cells(HSCs) are a kind of fat-storing cells, the lipid droplets are rich in the Cytoplasm, in which retinyl ester accounts for 42%, triglyceride occupies 28%, cholesterol (total) occupies 13%, pho...Hepatic stellate cells(HSCs) are a kind of fat-storing cells, the lipid droplets are rich in the Cytoplasm, in which retinyl ester accounts for 42%, triglyceride occupies 28%, cholesterol (total) occupies 13%, phospholipids occupies 4% respectively. Studies have continued that thetransforms of HSC phenotype follows the changing of the cell lipid. After the activation of HSC, with HSC phenotype changing from fat-storing cells into myofibroblast, the lipid droplets decreased or disappeared gradually, which means HSCs are under the differentiating process of removing adipose, meawhile triglyceride, and the main content of lipid droplets, also obviously reduced. It was ever declined that during the process of HSC re-fating, the activated HSC would turn into quiescent state. Therefore this shows HSCs fat metabolism is closely related to the biological activity.展开更多
Objective: To explore the possibility of the transfection of MyoD gene induced bone marrow mesenchymal stem cells ( MSCs) to differentiate into myoblasts in vitro. Methods: The eukaryotic expression plasmid vector pIR...Objective: To explore the possibility of the transfection of MyoD gene induced bone marrow mesenchymal stem cells ( MSCs) to differentiate into myoblasts in vitro. Methods: The eukaryotic expression plasmid vector pIRES2-EGFP-MyoD was transfected into MSCs with lipotransfection method, and the positive cells were selected by G418; The expression of MyoD was detected in the transfected MSCs with RT-PCR and the amplified, purified product was identified by sequencing; The reporter gene enhanced green fluorescence protein ( EFGP) was observed in the transfected cells under a fluorescent and a laser confocal microscopes; Immunohistochemical methods was used to examine the expressions of MyoD, myogenin, myosin, myoglobin and desmin in the differentiated cells. The ultrastructure changes of the cells before and after transfection were observed with electron microscopy. Results: The expression of MyoD was detected in the transfected MSCs with RT-PCR and the amplified, purified product was as same in sequence as that from Genbank; Green fluorescence was observed in the transfected cells under a fluorescent and a laser confocal microscopes; Immunohistochemical methods indicated that MyoD, myogenin, myosin, myoglobin and desmin were expressed in the transfected cells; The transfected cells showed the morphological characteristics of mature cells with filaments in their cytoplasm. Conclusion: MyoD gene can induce cultured MSCs to successfully differentiate into myoblasts , probably providing an experimental foundation for trauma repair.展开更多
文摘Hepatic stellate cells(HSCs) are a kind of fat-storing cells, the lipid droplets are rich in the Cytoplasm, in which retinyl ester accounts for 42%, triglyceride occupies 28%, cholesterol (total) occupies 13%, phospholipids occupies 4% respectively. Studies have continued that thetransforms of HSC phenotype follows the changing of the cell lipid. After the activation of HSC, with HSC phenotype changing from fat-storing cells into myofibroblast, the lipid droplets decreased or disappeared gradually, which means HSCs are under the differentiating process of removing adipose, meawhile triglyceride, and the main content of lipid droplets, also obviously reduced. It was ever declined that during the process of HSC re-fating, the activated HSC would turn into quiescent state. Therefore this shows HSCs fat metabolism is closely related to the biological activity.
文摘Objective: To explore the possibility of the transfection of MyoD gene induced bone marrow mesenchymal stem cells ( MSCs) to differentiate into myoblasts in vitro. Methods: The eukaryotic expression plasmid vector pIRES2-EGFP-MyoD was transfected into MSCs with lipotransfection method, and the positive cells were selected by G418; The expression of MyoD was detected in the transfected MSCs with RT-PCR and the amplified, purified product was identified by sequencing; The reporter gene enhanced green fluorescence protein ( EFGP) was observed in the transfected cells under a fluorescent and a laser confocal microscopes; Immunohistochemical methods was used to examine the expressions of MyoD, myogenin, myosin, myoglobin and desmin in the differentiated cells. The ultrastructure changes of the cells before and after transfection were observed with electron microscopy. Results: The expression of MyoD was detected in the transfected MSCs with RT-PCR and the amplified, purified product was as same in sequence as that from Genbank; Green fluorescence was observed in the transfected cells under a fluorescent and a laser confocal microscopes; Immunohistochemical methods indicated that MyoD, myogenin, myosin, myoglobin and desmin were expressed in the transfected cells; The transfected cells showed the morphological characteristics of mature cells with filaments in their cytoplasm. Conclusion: MyoD gene can induce cultured MSCs to successfully differentiate into myoblasts , probably providing an experimental foundation for trauma repair.