Phenotypic characterization of human smooth muscle cells derived from atherosclerotic tibial and peroneal arteries Bruce A. Jones, MD, Hamdy M. Aly,

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Presentation transcript:

Phenotypic characterization of human smooth muscle cells derived from atherosclerotic tibial and peroneal arteries Bruce A. Jones, MD, Hamdy M. Aly, MD, PhD, Elizabeth A. Forsyth, MD, Anton N. Sidawy, MD Journal of Vascular Surgery Volume 24, Issue 5, Pages 883-891 (November 1996) DOI: 10.1016/S0741-5214(96)70027-7 Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Composite photomicrograph demonstrating immunofluorescent images of monoclonal anti-α-smooth muscle actin antibody (passage 3). Extent of labeling to leading edge of cell is evident in A (open arrows). Most filaments are running along long axis of cells in B (solid arrow). Note also no nuclear staining. Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Composite photomicrograph demonstrating immunofluorescent images of monoclonal anti-α-smooth muscle actin antibody (passage 3). Extent of labeling to leading edge of cell is evident in A (open arrows). Most filaments are running along long axis of cells in B (solid arrow). Note also no nuclear staining. Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Composite electron micrograph of tibial smooth muscle cells in culture showing three phenotypes seen: A, contractile state (passage 3); B, intermediate state (passage 5); C, synthetic state (passage 7). In A, note extent of microfilaments (MF) throughout cytoplasm and highly irregular nuclear shape (N). Solid arrowheads denote dense plaques periodicity along MF. In B there is increase in number of ribosomes and polyribosomes (R). In later passage (C) there is disappearance of MF. LYS, Secondary lysosomes; LP, neural lipids. Magnification (A) ×4, 150, (B) ×3, 500, (C) ×5, 100. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Composite electron micrograph of tibial smooth muscle cells in culture showing three phenotypes seen: A, contractile state (passage 3); B, intermediate state (passage 5); C, synthetic state (passage 7). In A, note extent of microfilaments (MF) throughout cytoplasm and highly irregular nuclear shape (N). Solid arrowheads denote dense plaques periodicity along MF. In B there is increase in number of ribosomes and polyribosomes (R). In later passage (C) there is disappearance of MF. LYS, Secondary lysosomes; LP, neural lipids. Magnification (A) ×4, 150, (B) ×3, 500, (C) ×5, 100. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Composite electron micrograph of tibial smooth muscle cells in culture showing three phenotypes seen: A, contractile state (passage 3); B, intermediate state (passage 5); C, synthetic state (passage 7). In A, note extent of microfilaments (MF) throughout cytoplasm and highly irregular nuclear shape (N). Solid arrowheads denote dense plaques periodicity along MF. In B there is increase in number of ribosomes and polyribosomes (R). In later passage (C) there is disappearance of MF. LYS, Secondary lysosomes; LP, neural lipids. Magnification (A) ×4, 150, (B) ×3, 500, (C) ×5, 100. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Composite electron micrograph demonstrating portions of two synthetic tibial VSMCs (passage 7). A, Prominent Golgi apparatus (GA) and coated pits (solid arrowheads) can be seen. Extent of micropinocytotic vesicles can be seen in B, occurring either singularly (small solid arrows) or in clusters (open arrows). MV, Multivesicular bodies; CV, coated vesicles. Magnification, (A) ×32,000, (B) ×20,000. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Composite electron micrograph demonstrating portions of two synthetic tibial VSMCs (passage 7). A, Prominent Golgi apparatus (GA) and coated pits (solid arrowheads) can be seen. Extent of micropinocytotic vesicles can be seen in B, occurring either singularly (small solid arrows) or in clusters (open arrows). MV, Multivesicular bodies; CV, coated vesicles. Magnification, (A) ×32,000, (B) ×20,000. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Composite electron micrograph (passage 3 cells). A shows incomplete basal lamina (open arrows). Note close association of basal lamina with subplasmalemmal plaque (solid arrowheads) adjacent to where thin actin cortical filaments are ending (small solid arrows). B, Later passage (passage 7) shows almost complete basal lamina (open arrows). RER, Rough endoplasmic reticulum; DP, dense plaques; M, myelin figure. Magnification, (A) ×25,000, (B) ×16,000. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Composite electron micrograph (passage 3 cells). A shows incomplete basal lamina (open arrows). Note close association of basal lamina with subplasmalemmal plaque (solid arrowheads) adjacent to where thin actin cortical filaments are ending (small solid arrows). B, Later passage (passage 7) shows almost complete basal lamina (open arrows). RER, Rough endoplasmic reticulum; DP, dense plaques; M, myelin figure. Magnification, (A) ×25,000, (B) ×16,000. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Composite electron micrograph showing extensive extracellular basal lamina-like matrix deep to multilayered tibial smooth muscle cells in culture (A, open arrows). There is also thickened basal laminae (open arrows) associated with smooth muscle cells from atherosclerotic tibial artery as indicated in B. MF, Microfilaments; M, mitochondria; GA, Golgi apparatus; RER, rough endoplasmic reticulum; N, nucleus; DP, dense plaques, arrows, cluster of micro-pinocytotic vesicles. Magnification, (A) ×12,500, (B) ×8,500. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Composite electron micrograph showing extensive extracellular basal lamina-like matrix deep to multilayered tibial smooth muscle cells in culture (A, open arrows). There is also thickened basal laminae (open arrows) associated with smooth muscle cells from atherosclerotic tibial artery as indicated in B. MF, Microfilaments; M, mitochondria; GA, Golgi apparatus; RER, rough endoplasmic reticulum; N, nucleus; DP, dense plaques, arrows, cluster of micro-pinocytotic vesicles. Magnification, (A) ×12,500, (B) ×8,500. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Composite phase contrast micrograph of tibial smooth muscle cells cultured on rubber substratum (passage 3). A, Before adding angiotensin II. B, Response after adding angiotensin II (3 ×10-7 mol/L, solid arrowheads). C, Cells seen before adding endothelin I. D, Twenty minutes after adding endothelin I (10-7 mol/L), a marked increase in wrinkling and lengthening of preexisting wrinkles are seen (open arrows). E, Cells seen before adding bradykinin. F, Contraction response seen after addition of bradykinin (3 × 10-7 mol/L, solid arrows). Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Composite phase contrast micrograph of tibial smooth muscle cells cultured on rubber substratum (passage 3). A, Before adding angiotensin II. B, Response after adding angiotensin II (3 ×10-7 mol/L, solid arrowheads). C, Cells seen before adding endothelin I. D, Twenty minutes after adding endothelin I (10-7 mol/L), a marked increase in wrinkling and lengthening of preexisting wrinkles are seen (open arrows). E, Cells seen before adding bradykinin. F, Contraction response seen after addition of bradykinin (3 × 10-7 mol/L, solid arrows). Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Composite phase contrast micrograph of tibial smooth muscle cells cultured on rubber substratum (passage 3). A, Before adding angiotensin II. B, Response after adding angiotensin II (3 ×10-7 mol/L, solid arrowheads). C, Cells seen before adding endothelin I. D, Twenty minutes after adding endothelin I (10-7 mol/L), a marked increase in wrinkling and lengthening of preexisting wrinkles are seen (open arrows). E, Cells seen before adding bradykinin. F, Contraction response seen after addition of bradykinin (3 × 10-7 mol/L, solid arrows). Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Composite phase contrast micrograph of tibial smooth muscle cells cultured on rubber substratum (passage 3). A, Before adding angiotensin II. B, Response after adding angiotensin II (3 ×10-7 mol/L, solid arrowheads). C, Cells seen before adding endothelin I. D, Twenty minutes after adding endothelin I (10-7 mol/L), a marked increase in wrinkling and lengthening of preexisting wrinkles are seen (open arrows). E, Cells seen before adding bradykinin. F, Contraction response seen after addition of bradykinin (3 × 10-7 mol/L, solid arrows). Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Composite phase contrast micrograph of tibial smooth muscle cells cultured on rubber substratum (passage 3). A, Before adding angiotensin II. B, Response after adding angiotensin II (3 ×10-7 mol/L, solid arrowheads). C, Cells seen before adding endothelin I. D, Twenty minutes after adding endothelin I (10-7 mol/L), a marked increase in wrinkling and lengthening of preexisting wrinkles are seen (open arrows). E, Cells seen before adding bradykinin. F, Contraction response seen after addition of bradykinin (3 × 10-7 mol/L, solid arrows). Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Composite phase contrast micrograph of tibial smooth muscle cells cultured on rubber substratum (passage 3). A, Before adding angiotensin II. B, Response after adding angiotensin II (3 ×10-7 mol/L, solid arrowheads). C, Cells seen before adding endothelin I. D, Twenty minutes after adding endothelin I (10-7 mol/L), a marked increase in wrinkling and lengthening of preexisting wrinkles are seen (open arrows). E, Cells seen before adding bradykinin. F, Contraction response seen after addition of bradykinin (3 × 10-7 mol/L, solid arrows). Magnification ×200. Journal of Vascular Surgery 1996 24, 883-891DOI: (10.1016/S0741-5214(96)70027-7) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions