Platelet-derived growth factor and extracellular matrix proteins provide a synergistic stimulus for human vascular smooth muscle cell migration Peter.

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Presentation transcript:

Platelet-derived growth factor and extracellular matrix proteins provide a synergistic stimulus for human vascular smooth muscle cell migration Peter R. Nelson, MD, Shinji Yamamura, MD, K.Craig Kent, MD Journal of Vascular Surgery Volume 26, Issue 1, Pages 104-112 (July 1997) DOI: 10.1016/S0741-5214(97)70153-8 Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Smooth muscle cell (SMC) chemotaxis in response to combinations of platelet-derived growth factor (PDGF) and extracellular matrix (ECM) proteins. SMC chemotaxis was measured in response to PDGF: laminin (LM; A), fibronectin (FN; B), collagen I (CN I; C), collagen IV (CN IV; D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMCs simultaneously with PDGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Smooth muscle cell (SMC) chemotaxis in response to combinations of platelet-derived growth factor (PDGF) and extracellular matrix (ECM) proteins. SMC chemotaxis was measured in response to PDGF: laminin (LM; A), fibronectin (FN; B), collagen I (CN I; C), collagen IV (CN IV; D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMCs simultaneously with PDGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Smooth muscle cell (SMC) chemotaxis in response to combinations of platelet-derived growth factor (PDGF) and extracellular matrix (ECM) proteins. SMC chemotaxis was measured in response to PDGF: laminin (LM; A), fibronectin (FN; B), collagen I (CN I; C), collagen IV (CN IV; D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMCs simultaneously with PDGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Smooth muscle cell (SMC) chemotaxis in response to combinations of platelet-derived growth factor (PDGF) and extracellular matrix (ECM) proteins. SMC chemotaxis was measured in response to PDGF: laminin (LM; A), fibronectin (FN; B), collagen I (CN I; C), collagen IV (CN IV; D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMCs simultaneously with PDGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 SMC chemotaxis in response to combinations of epidermal growth factor (EGF) and (ECM) proteins. SMC chemotaxis was measured in response to EGF: LM (A), FN (B), CN I (C), CN IV (D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMC simultaneously with EGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 SMC chemotaxis in response to combinations of epidermal growth factor (EGF) and (ECM) proteins. SMC chemotaxis was measured in response to EGF: LM (A), FN (B), CN I (C), CN IV (D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMC simultaneously with EGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 SMC chemotaxis in response to combinations of epidermal growth factor (EGF) and (ECM) proteins. SMC chemotaxis was measured in response to EGF: LM (A), FN (B), CN I (C), CN IV (D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMC simultaneously with EGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 SMC chemotaxis in response to combinations of epidermal growth factor (EGF) and (ECM) proteins. SMC chemotaxis was measured in response to EGF: LM (A), FN (B), CN I (C), CN IV (D), or combinations of these factors in the concentrations indicated. The calculated additive effect, derived as described in the Material and Methods section from the individual responses (open bars), is compared with chemotaxis that results from coincubating SMC simultaneously with EGF and ECM proteins (solid bars). Results are expressed as fold increase ± SD compared with migration of unstimulated control. Experiments were performed in triplicate and repeated with use of at least three cell lines from different donors. Data from a representative experiment are shown. *p < 0.05, coincubation versus calculated additive effect. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Measurement of intracellular calcium ([Ca2+]i) in response to PDGF and ECM proteins. Quiescent human SMCs were loaded with 2 μmol/L fura-2-acetoxymethyl ester and stimulated with either PDGF-AB (5 ng/ml) alone (broken lines), ECM proteins alone (dotted lines) [LM (A), FN (B), CN I (C), or CN IV (D)] or with combinations of PDGF and ECM proteins (solid lines). [Ca2+]i was detected by fluorimetry as described in the Material and Methods section. Experiments were performed in at least three cell lines from different donors. Curves from a representative experiment are shown. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Measurement of intracellular calcium ([Ca2+]i) in response to PDGF and ECM proteins. Quiescent human SMCs were loaded with 2 μmol/L fura-2-acetoxymethyl ester and stimulated with either PDGF-AB (5 ng/ml) alone (broken lines), ECM proteins alone (dotted lines) [LM (A), FN (B), CN I (C), or CN IV (D)] or with combinations of PDGF and ECM proteins (solid lines). [Ca2+]i was detected by fluorimetry as described in the Material and Methods section. Experiments were performed in at least three cell lines from different donors. Curves from a representative experiment are shown. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Measurement of intracellular calcium ([Ca2+]i) in response to PDGF and ECM proteins. Quiescent human SMCs were loaded with 2 μmol/L fura-2-acetoxymethyl ester and stimulated with either PDGF-AB (5 ng/ml) alone (broken lines), ECM proteins alone (dotted lines) [LM (A), FN (B), CN I (C), or CN IV (D)] or with combinations of PDGF and ECM proteins (solid lines). [Ca2+]i was detected by fluorimetry as described in the Material and Methods section. Experiments were performed in at least three cell lines from different donors. Curves from a representative experiment are shown. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Measurement of intracellular calcium ([Ca2+]i) in response to PDGF and ECM proteins. Quiescent human SMCs were loaded with 2 μmol/L fura-2-acetoxymethyl ester and stimulated with either PDGF-AB (5 ng/ml) alone (broken lines), ECM proteins alone (dotted lines) [LM (A), FN (B), CN I (C), or CN IV (D)] or with combinations of PDGF and ECM proteins (solid lines). [Ca2+]i was detected by fluorimetry as described in the Material and Methods section. Experiments were performed in at least three cell lines from different donors. Curves from a representative experiment are shown. Journal of Vascular Surgery 1997 26, 104-112DOI: (10.1016/S0741-5214(97)70153-8) Copyright © 1997 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions