Acta Anatomica Sinica ›› 2017, Vol. 48 ›› Issue (1): 43-47.doi: 10.16098/j.issn.0529-1356.2017.01.008

• Cell and Molecules Biology • Previous Articles     Next Articles

Cardiac fibroblasts of the rat transfected by lenti virus-stromal cell-derived factor-1α-green fluorescent protein vector

ZHU Zhan-zhan1 HOU Bin2 SUN Wen-wen3 YUAN Hui-fang1 ZHANG Yong-chun4 CAI Xin-hua 1*   

  1. 1. Key Laboratory for Tissue Regeneration of He’nan Province,Xinxiang Medical University, He’nan Xinxiang 453003, China; 2. Department of Cardiology, Pingdingshan Second People’s Hospital, He’nan Pingdingshan 467000, China; 3. Department of Infectious Diseases, 4. Department of Cardiology, the First Affiliated Hospital of Xinxiang Medical College, He’nan Weihui 453003, China

  • Received:2016-06-14 Revised:2016-09-28 Online:2017-02-06 Published:2017-02-06
  • Contact: CAI Xin-hua E-mail:cxh@xxmu.edu.cn

Abstract:

Objective To investigate the effect of lentivirus-stromal cell-derived factor-1α-green fluorescent protein(LV-SDF-1α-GFP) on the cardiac fibroblasts, the optimum conditions of infection, the expression and secretion of the target protein. Methods The cardiac fibroblasts of neonatal rats were primarily isolated and cultured by differential adherence methods, and were observed and identifi with immunofluorescence. LV-SDF-1α-GFP with different titers and conditions was transfected into cardiac fibroblasts. The expression of fluorescence and the optimal transfection conditions were observed. LV-SDF-1α-GFP target gene virus and negative control CON145 virus were transfected into cardiac fibroblasts. The growth curve was drawn, and the effect of transfection on the proliferation of cardiac fibroblasts was explored.The cardiac fibroblasts were transfected with the optimum transfection dose, and the expression of SDF-1α was detected by Dot-blotting. The measurement data underwent statistical analysis. Results There was no statistical difference between the cardiac fibroblasts with SDF-1α transfected lentivirus and without no-transfected SDF-1α lentivirus. The peak of the expression of SDF-1α appeared in culture day 4 and statistical analysis showed significantly difference (P<0.05). Conclusion The LV-SDF-1α-GFP vector is of higher transfection efficiency to cardiac fibroblasts with the both low cytotoxicity and ability of secreting SDF-1α protein.

Key words: Cardiac fibroblast, Lentivirus-stromal cell-derived factor-1α-green fluorescent protein vector, Transfection, Dot blotting, Neonatal rat