Presentation is loading. Please wait.

Presentation is loading. Please wait.

Retroviral-mediated transduction of endothelial cells with the lac Z gene impairs cellular proliferation in vitro and graft endothelialization in vivo 

Published byLeszek Sawicki Modified over 5 years ago

Similar presentations

Presentation on theme: "Retroviral-mediated transduction of endothelial cells with the lac Z gene impairs cellular proliferation in vitro and graft endothelialization in vivo "— Presentation transcript:

1 Retroviral-mediated transduction of endothelial cells with the lac Z gene impairs cellular proliferation in vitro and graft endothelialization in vivo  Rachel Podrazik Baer, MD, Thomas E. Whitehill, MD, Rajabrata Sarkar, MD, Minakshi Sarkar, BS, Louis M. Messina, MD, Thomas A. Komorowski, MS, James C. Stanley, MD  Journal of Vascular Surgery  Volume 24, Issue 5, Pages (November 1996) DOI: /S (96) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

2 Fig. 1 Proliferation of transduced and nontransduced ECs. Proliferation of late-passage ECs was less than that of early-passage ECs at all times (*p < 0.05, ANOVA, Scheffe's test), except late-passage nontransduced ECs achieved similar confluent densities as early-passage ECs by 14 days. Among early-passage ECs proliferation of lac Z-transduced ECs was significantly less at days 3 and 5 (**p < 0.05, ANOVA). Among late-passage cells proliferation of lac Z-transduced ECs was significantly less than neomycin resistance-transduced ECs and nontransduced ECs at days 3, 5, and 9 (***p < 0.05, ANOVA). Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

3 Fig. 1 Proliferation of transduced and nontransduced ECs. Proliferation of late-passage ECs was less than that of early-passage ECs at all times (*p < 0.05, ANOVA, Scheffe's test), except late-passage nontransduced ECs achieved similar confluent densities as early-passage ECs by 14 days. Among early-passage ECs proliferation of lac Z-transduced ECs was significantly less at days 3 and 5 (**p < 0.05, ANOVA). Among late-passage cells proliferation of lac Z-transduced ECs was significantly less than neomycin resistance-transduced ECs and nontransduced ECs at days 3, 5, and 9 (***p < 0.05, ANOVA). Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

4 Fig. 2 En face X-gal-stained graft surfaces (×8). Panel on right shows region of β-galactosidase-expressing ECs from lac Z-transduced EC seeded graft. Among transduced seeded grafts such foci of blue-stained ECs were distributed throughout surface of graft. Panel on left shows nontransduced EC seeded graft surface without evidence of X-gal β-galactosidase cellular staining. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

5 Fig. 3 Cross-sections of grafts (presented in Fig. 2) after X-gal staining for β-galactosidase and counterstaining with hematoxylin-eosin (×120). Blue-stained EC nomolayer is evident on surface of lac Z-transduced EC seeded graft (right panel). No blue staining is seen on nontransduced EC seeded graft (left). Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

6 Fig. 4 Mean percent total graft surface endothelialization for each graft type and mean percent of graft surface area that demonstrated ECs with β-galactosidase expression. Spontaneous coverage by ingrowth of native endothelium was documented in nonseeded grafts at 31%. Among three groups of EC seeded grafts, nontransduced ECs achieved 79% coverage, significantly greater than all other graft types (*p < 0.05, ANOVA, Scheffe's test). Lac Z-transduced EC seeded grafts are divided into two groups for analysis: those grafts seeded with early-passage ECs (passage 2 to 4) achieved 28% total surface EC coverage, and those seeded with late-passage ECs (passage 6 to 9) achieved 42% total surface EC coverage. Less than half of endothelialized areas in grafts seeded with lac Z-transduced ECs were covered by β-galactosidase expressing ECs. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

7 Fig. 5 Representative gel after DNA PCR of sampled luminal cellular surface material from endothelialized areas of three lac Z-transduced EC seeded grafts that did not exhibit lac Z gene expression by X-gal staining. No lac Z DNA is present in graft specimens 14-20, 17-18, and 18-12, in contrast with expected lac Z EC DNA from positive X-gal staining region of graft specimen 18-7. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

8 Fig. 6 Nonspecific blue staining with X-gal reagent (x8). In panel on left, region from nontransduced EC seeded graft that was stained with appropriate technique reveals pattern of noncellular light blue staining. Panel on right represents segment of nontransduced EC seeded graft that was purposely overstained. These examples demonstrate potential for error when relying on blue-stained appearance alone as evidence of lac Z gene expression. Journal of Vascular Surgery  , DOI: ( /S (96) ) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Download ppt "Retroviral-mediated transduction of endothelial cells with the lac Z gene impairs cellular proliferation in vitro and graft endothelialization in vivo "

Similar presentations

Endothelial cell seeding reduces thrombogenicity of Dacron grafts in humans Per Örtenwall, MD, PhD *, Hans Wadenvik, MD, PhD **, Jack Kutti, MD, PhD **,

Endothelial cell seeding reduces thrombogenicity of Dacron grafts in humans Per Örtenwall, MD, PhD *, Hans Wadenvik, MD, PhD **, Jack Kutti, MD, PhD **,
The effect of inguinal lymphatic manipulation on regional lymph flow patterns Jeffrey R. Rubin, MD, Lisa B. Eberlin, MD Journal of Vascular Surgery Volume.

The effect of inguinal lymphatic manipulation on regional lymph flow patterns Jeffrey R. Rubin, MD, Lisa B. Eberlin, MD Journal of Vascular Surgery Volume.
Shunya Shindo, M. D. , Atsuhiko Takagi, M. D. , Anthony D

Shunya Shindo, M. D. , Atsuhiko Takagi, M. D. , Anthony D
In vitro endothelialization of a mesosystemic shunt: A clinical case report  Teddy Fischlein, MD, Peter Zilla, MD, PhD, Johann Meinhart, PhD, Rudolf Puschmann,

In vitro endothelialization of a mesosystemic shunt: A clinical case report  Teddy Fischlein, MD, Peter Zilla, MD, PhD, Johann Meinhart, PhD, Rudolf Puschmann,
Axillary-to-carotid artery bypass grafting for symptomatic severe common carotid artery occlusive disease  Joseph P. Archie, PhD, MD  Journal of Vascular.

Axillary-to-carotid artery bypass grafting for symptomatic severe common carotid artery occlusive disease  Joseph P. Archie, PhD, MD  Journal of Vascular.
Gastroesophageal tumor embolization to the popliteal arteries

Gastroesophageal tumor embolization to the popliteal arteries
Adenoviral-mediated expression of antisense RNA to basic fibroblast growth factor reduces tangential stress in arterialized vein grafts  Abigail K. Hanna,

Adenoviral-mediated expression of antisense RNA to basic fibroblast growth factor reduces tangential stress in arterialized vein grafts  Abigail K. Hanna,
Gene transfer to coronary artery bypass conduits

Gene transfer to coronary artery bypass conduits
Preformed confluent endothelial cell monolayers prevent early platelet deposition on vascular prostheses in baboons  Peter A. Schneider, MD, Stephen R.

Preformed confluent endothelial cell monolayers prevent early platelet deposition on vascular prostheses in baboons  Peter A. Schneider, MD, Stephen R.
Somatostatin gene transfer and expression in endothelial cells

Somatostatin gene transfer and expression in endothelial cells
The differential effect of contrast agents on endothelial cell and smooth muscle cell growth in vitro  Carol J. Sawmiller, MD, Richard J. Powell, MD,

The differential effect of contrast agents on endothelial cell and smooth muscle cell growth in vitro  Carol J. Sawmiller, MD, Richard J. Powell, MD,
Mark Wengrovitz, MD. , Lulseged G. Selassie, MD. , Robert R. M

Mark Wengrovitz, MD. , Lulseged G. Selassie, MD. , Robert R. M
Vida K. Stark, MS, Thomas F. Warner, MD, John R. Hoch, MD 

Vida K. Stark, MS, Thomas F. Warner, MD, John R. Hoch, MD 
A new valvulotome for in situ bypass grafts

A new valvulotome for in situ bypass grafts
Thomas E. Arnold, MD, Dmitri Gnatenko, PhD, Wadie F. Bahou, MD 

Thomas E. Arnold, MD, Dmitri Gnatenko, PhD, Wadie F. Bahou, MD 
Makoto Hashizume, M. D. , Ph. D. , Yeng Yang, M. D. , Spencer Galt, M

Makoto Hashizume, M. D. , Ph. D. , Yeng Yang, M. D. , Spencer Galt, M
Chronic in vitro shear stress stimulates endothelial cell retention on prosthetic vascular grafts and reduces subsequent in vivo neointimal thickness 

Chronic in vitro shear stress stimulates endothelial cell retention on prosthetic vascular grafts and reduces subsequent in vivo neointimal thickness 
Mark F. Fillinger, MD, Susan E. O'Connor, MD, Robert J

Mark F. Fillinger, MD, Susan E. O'Connor, MD, Robert J
Proof of fallout endothelialization of impervious Dacron grafts in the aorta and inferior vena cava of the dog  Qun Shi, MD, Moses Hong-De Wu, MD, Naoki.

Proof of fallout endothelialization of impervious Dacron grafts in the aorta and inferior vena cava of the dog  Qun Shi, MD, Moses Hong-De Wu, MD, Naoki.
Improved retroviral transduction efficiency of vascular cells in vitro and in vivo during clinically relevant incubation periods using centrifugation.

Improved retroviral transduction efficiency of vascular cells in vitro and in vivo during clinically relevant incubation periods using centrifugation.

Similar presentations

Ads by Google