Somatostatin gene transfer and expression in endothelial cells

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Presentation transcript:

Somatostatin gene transfer and expression in endothelial cells Rajabrata Sarkar, MD, PhD, Susan Finniss, MS, Chris J. Dickinson, MD, James C. Stanley, MD Journal of Vascular Surgery Volume 27, Issue 5, Pages 955-962 (May 1998) DOI: 10.1016/S0741-5214(98)70278-2 Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Retroviral vectors used to transfect ECs. BAG vector (top) with bacterial b-galactosidase gene driven by retroviral long terminal repeat (LTR) promoter. Neomycin phosphotransferase gene (neomycin) is driven by the Simian virus 40 (SV40) promoter. Human somatostatin vector with the human somatostatin cDNA (somatostatin) in place of β-galactosidase gene. Journal of Vascular Surgery 1998 27, 955-962DOI: (10.1016/S0741-5214(98)70278-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Analysis of somatostatin peptide produced by transfected CVEC. A, Chromatographic separation of somatostatin-14 and somatostatin-28 peptide standards followed by somatostatin immunoreactivity assay. B, Analysis of HSS EC medium under same conditions. Journal of Vascular Surgery 1998 27, 955-962DOI: (10.1016/S0741-5214(98)70278-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Somatostatin production by BAG- and HSS-transfected CVECs. Transfected CVECs were grown to confluence, and the medium was replaced and collected after 24 hours for somatostatin immunoreactivity determination (n = 3, *p < 0.05 vs BAG group). Journal of Vascular Surgery 1998 27, 955-962DOI: (10.1016/S0741-5214(98)70278-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Growth of CVECs over 96 hours after transfection with BAG or HSS retrovirus. After retroviral transfection and selection, cells were plated at a concentration of 5,000 cells/cm2, and the increase in cell number over 96 hours was determined (n = 4 per group). Journal of Vascular Surgery 1998 27, 955-962DOI: (10.1016/S0741-5214(98)70278-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Somatostatin production by various EC types after BAG and HSS transfection and selection. Transfected ECs were grown to confluence, and the medium was changed and collected after 24 hours for somatostatin immunoreactivity determination (n = 3, *p < 0.05 vs all other groups). Journal of Vascular Surgery 1998 27, 955-962DOI: (10.1016/S0741-5214(98)70278-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Microscopic analysis of heterogeneity of β-galactosidase expression in endothelium after BAG transfection and selection. A, Canine venous endothelium (CVECs). B, Rat aortic endothelium (RAECs). C, Rat microvascular endothelium (RMVECs). Shown are cells expressing β-galactosidase (arrow) or not expressing β-galactosidase (arrowhead). Original magnification, ×100. Journal of Vascular Surgery 1998 27, 955-962DOI: (10.1016/S0741-5214(98)70278-2) Copyright © 1998 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions