title: [A experimental study on transfecting human stromal cell-derived factor 1alpha and human vascular endothelial growth factor 165 genes into myoblasts in vitro].
abstract: OBJECTIVE
  To explore the human stromal cell-derived factor 1alpha (hSDF-1alpha) and human vascular endothelial growth factor 165 (hVEGF165) mRNA expressions of the transfected cells after hSDF-1alpha gene and hVEGF165 gene were transfected into rat myoblasts in vitro so as to lay a foundation for further study on the synergistic effects of 2 genes on tissue engineered skeletal muscle vascularization.
  
  
  METHODS
  The myoblasts of 1-day-old Sprague Dawley rats were cultured and purified by trypsin digestion assay in vitro and were identified by immunohistochemistry staining of Desmin. Approximately 70%-80% of confluent myoblasts were transfected with enhanced green fluorescent protein (EGFP)-hSDF-1alpha and EGFP-hVEGF165 genes in vitro (transfected group) and were not transfected (control group). The expressions of hSDF-1alpha and hVEGF165 mRNA and protein in the transfected cells were detected by RT-PCR, ELISA, and Western blot respectively.
  
  
  RESULTS
  The cultured cells were identified as myoblasts by immunohistochemistry staining of Desmin. The expression of green fluorescent protein was observed in transfected cells, indicating that hSDF-1alpha and hVEGF165 genes were transfected into myoblasts successfully. The mRNA and protein expressions of the 2 genes were positive in the transfected group by RT-PCR and Western bolt assay at 2, 4, 6, and 8 days after transfection, and were negative in the control group. The expressions of hSDF-1alpha and hVEGF165 showed a stable low level in the control group, but the expressions of the proteins increased at 2 days and then showed gradual downtrend with time in the transfected group by ELISA assay. There were significant differences in the expressions of hSDF-1alpha and hVEGF165 proteins between different time points in the transfected group, and between 2 groups (P < 0.05).
  
  
  CONCLUSIONS
  hSDF-1alpha and hVEGF165 genes are successfully transfected into myoblasts in vitro, and mRNA and proteins of hSDF-1alpha and hVEGF165 can be expressed in the transfected myoblasts, which may provide the experimental evidence for the expressions of hSDF-1alpha and hVEGF165 mRNA and proteins in vivo successfully.
authors: Department of Stomatology, First Hospital of Lanzhou University, Lanzhou Gansu 730000, PRChina.
authors: Dengqi
authors: D
authors: He
authors: Xiufa
authors: X
authors: Tang
authors: Chengge
authors: C
authors: Hua
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000235:genetics*; D006801:Humans; D008297:Male; D032446:Myoblasts / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*
chemical_list: D054377:Chemokine CXCL12; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A