Abstract:

BACKGROUND: Recently, application of stem cells and growth factor to promoting lung regeneration in repair of emphysema lesion has been a hot focus in study. Thus, it is worth to pay attention on whether stem cells carrying relevant foreign growth factor gene can repair emphysema lesion.
OBJECTIVE: To evaluate the efficiency of adenovirus vector mediated green fluorescence protein (Ad-GFP) transfecting bone marrow mesenchymal stem cells (BMSCs) and its effect on the cell proliferation, to explore oriented migration of intravenously administrated BMSCs transfected with Ad-GFP in the lung tissues of pulmonary emphysema rats.
METHODS: MSCs were separated and purified from the bone marrow of rats by density gradient centrifugation and by adherence. At different multiplicity of infection (MOI), transfection efficiency was observed by laser confocal microscopy. At 48 hours of transfection, MTT method was used to evaluate the proliferation of MSCs. A total of 16 Wistar rats were randomly divided into emphysema model group and control group (n = 8). Model rats were established by exposure to cigarette smoke. MSCs, transfected with Ad-GFP, were grafted into the body of rats via tail vein. Lungs derived at 24 hours after implantation, and frozen sections were made. Migration and survival of MSCs in the lung tissues were observed by fluorescence microscopy.
RESULTS AND CONCLUSION: MSCs from Wistar rats were successfully cultured, grew well and infected by Ad-GFP. The highest transfection effincincy (88.42 %) could be achieved at MOI of 200. Green fluorescent protein labeling had little effect on proliferation of MSCs by different MOI (P > 0. 05). At 24 hours posttransplantation, the green fluorescence-positive tissue was found in the lung tissues of emphysema model group and control group. Compared with control group, the expression of GFP in lung tissues was higher in emphysema model group (P < 0. 05). These suggested that introduction of target gene cannot affect proliferation and homing property of BMSCs.