Vascular smooth muscle cells cell culture

Grainger DJ, Weissberg PL, Metcalfe JC (1993) Tamoxifen decreases the rate of proliferation of rat vascular smooth-muscle cells in culture by inducing production of transforming growth factor beta. Biochem J 294 109-112... 

Figure 7. Talin-containing streak-like focal adhesion plaques (A) and beta-actin cytoskeleton (B) in vascular smooth muscle cells in cultures derived from the rat aorta. Immunofluorescence staining, the nuclei couterstained with propidium iodide (A) or Hoechst 33342 (B). Microscope Olympus IX 50, digital camera DP 70, obj. 100, bar=20 pm.

Buemi M, etal. Effects of homocysteine on proliferation, necrosis, and apoptosis of vascular smooth muscle cells in culture and influence of folic acid. Thromb Res 2001 104(3) 207-213. 

Ortego Monica, et al. 3-Hydroxy-3-methylglutaryl coenzyme A reductase and isoprenylation inhibitors induce apoptosis of vascular smooth muscle cells in culture. Circ. Res., 1998, 83, 490-500. 

By copolymerizing the monomers of two highly crystalline homopolymers it is possible to create a series of less crystalline copolymers with reduced levels of stiffness, elastic modulus and crystallinity depending on the blend level of the two monomers. This has been demonstrated by Soo H5nm Kim and coworkers at KIST, who have been able to synthesize a series of elastomeric (or rubbery) resorbable poly(L-lactide-co-e-caprolactone) copolymers. Because of their superior elastic behavior they have been used to prepare tubular scaffolds for vascular smooth muscle cell (SMC) culture under dynamic pulsatile conditions. We have melt spun and electrospun the 50/50 copolymer to fabricate a two-layer small-caliber vascular prosthesis prototype with radial elongation at break in excess of 300% (Fig. 15.33). " ... 

Bacakova L, Filova E, Kubies D, Machova L, Proks V, Malinova V, Lisa V, Rypacek F. Adhesion and growth of vascular smooth muscle cells in cultures on bioactive RDG peptide-carrying polylactides. / Mater. Sci. Mater. Med. 18 ... 

Bacakova L, Mares V, Grazia Bottone M, Pellicciari C, Lisa V, Svorcik V. Fluorine ion-implanted polystyrene improves growth and viability of vascular smooth muscle cells in culture. J Biomed Mater Res 2000 49 369-79. 

Another example was done by Opitz et al. They utilized P4HB scaffolds to produce viable ovine blood vessels, and then implanted the blood vessels in the systemic circulation of sheep. Enzymatically derived vascular smooth muscle cells (vSMC) were seeded on the scaffolds both under pulsatile flow and static conditions. Mechanical properties of bioreactor-cultured blood vessels which were obtained from tissue engineering approached those of native aorta. 

Frackowiak J, Mazur Kolecka B, Wisniewski HM, Potempska A, Carroll RT, Emmerling MR, Kim KS. Secretion and accumulation of Alzheimer s beta-protein by cultured vascular smooth muscle cells from old and young dogs. Brain Res 1995 676 225-230. 

Cultured rat vascular smooth muscle cells (VSMCs), grown and prepared for respirometry as described in Doeller et al., 2005 [41], were injected into the respirometer chamber to a concentration of between 105 and 106 cells ml 1. Cell viability remained at >90% throughout experiments. Near 4pM 02, H2S production was stimulated by the addition of L-cysteine and PLP (Fig. 8.8). The initial H2S production rate was approximately 20% of the rat aorta homogenate rate. H2S production rate decreased after the initial rise in H2S concentration, perhaps the result of product feedback inhibition. The addition of the CGL inhibitor BCA showed an effect similar to aorta homogenate. 

FIGURE 8.8 H2S production in vascular tissues. IPS production by aorta homogenate (upper panel), cultured rat vascular smooth muscle cells (VSMCs middle panel), and intact rat aorta occurs after the addition of substrate L-cysteine (L-cys) and cofactor pyridoxal L-phosphate (PLP) for the enzyme CGL located in vascular tissue. H2S production is inhibited after the CGL. 3 cyano-L-alanine (BCA) is added. Ferric Lucina pectinata hemoglobin I (metHb) is added to confirm H2S production. The quantity of metHb-sulfide produced, determined spectrophotometrically, matched the levels of H2S detected by the PHSS (after [41]). 

Govindarajan G, EbleDM, Lucchesi PA, Samarel AM 2000 Focal adhesion kinase is involved in angiotensin Il-mediated protein synthesis in cultured vascular smooth muscle cells. Circ Res... 

Nakamura M, Sunagawa M, Kosugi T, Sperelakis N 2000 Actin filament disruption inhibits L-type Ca2+ channel current in cultured vascular smooth muscle cells. Am J Physiol 279 0480-0487... 

Figure 22. Human embryonic kidney cells (A), rat vascular smooth muscle cells (B, C) and human osteoblast-like MG 63 cells (D) in cultures on micropattemed surfaces. A, B PTFE irradiated with UV light produced by a Xe2 -excimer lamp for 30 min in an ammonia atmosphere through a mask with holes 100 pm in diameter and center-to-center distance 300 pm C PE irradiated with Ar ions (energy 150 keV, ion dose lO ions/cm ) through a mask with holes 100 pm in diameter and center-to-center distance 200 pm fullerenes Qo deposited through a mask with rectangular holes with an average size of 128 3 pm per 98 8 pm on glass coverslips. Day 7 after seeding. A native cells in an inverted phase-contrast microscope B, C cells stained with hematoxylin and eosin, Olympus microscope IX 50 D cells stained with fluorescence-based LIVE/DEAD viability/cytotoxicity kit (Invitrogen), Olympus microscope IX 50. Bars 300 pm (A), 200 pm (B, D), Imm (C) [10,11].

Figure 25. A-D Immunofluorescence staining of vmculrn in vascular smooth muscle cells on day 3 after seeding on polymeric surfaces (medium supplemented with 10% fetal bovine serum). A poly(DL-lactic acid), PDLLA B block copolymer of poly(DL-lactic acid) and poly (ethylene oxide) (PEO), PDLLA-6-PEO C, E PDLLA-6-PEO with 5% GRGDSG-PEO-6-PDLLA D, F PDLLA-6-PEO with 20% GRGDSG-PEO-6-PDLLA. E, F Immunoperoxidase staining of bromodeoxyuridine (arrows) incorporated into DNA newly synthesized in vascular smooth muscle cells cultured for 3 days in serum-free medium on PDLLA-Z)-PEO with 5% (E) or 20% (F) GRGDSG-PEO-6-PDLLA. Cells counterstained with light green. Bar=100 pm [41].

Matsui T, Ueno T, Tanaka M, Oka H, Miyamoto T, Osajima K, Matsumoto K. (2005) Antiproliferation action of an angiotensin I-converting enzyme inhibitory peptide, Val-Tyr, via an L-type Ca channel inhibition in cultured vascular smooth muscle cells. Hypertens Res 28 545-552. 

Garg, U. C., and Hassid, A. (1989). Nitric oxide-generating vasodilators and 8-bromo-cyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells. J. Clin. Invest. 83, 1774-1777. 

Rennick RE, Connat JL, Burnstock G, Rothery S, Severs NJ, Green CR Expression of connexin 43 gap junctions between cultured vascular smooth muscle cells is dependent upon phenotype. Cell Tissue Res 1993 271 323-332. 

DNA binding moiety has been synthesized and evaluated as a receptor-mediated gene delivery system. The system was able to transfect about 30% of corneal endothelial cells of rabbit, pig and human in the presence of chloroquine (Shewring et al., 1997). Associated with the fusogenic peptide of influenza, this peptide transfected 25-30% of primary cultures of vascular smooth muscle cells of man, rabbit and rat (Collins et al., 2000 Li et al., 2000). The molossin-based gene delivery system represents an interesting system in transplantation because the molossin peptide does not bind to vascular endothelium and pancreatic islets. 

Patel, M. K., Churn, G. F., Lymn, J. S., Austin, O. and Hughes, A. D., 2005, Effect of serum withdrawal on die contribution of L-type calcium channels (CaV1.2) to intracellular Ca2+ responses and chemotaxis in cultured human vascular smooth muscle cells. Br J Pharmacol 145, 811—7. 

Increases in plasma S-AA levels have previously been reported in patients with coronary disease (57). S-AA and plasma intracellular adhesion molecule-1 were elevated in patients with CAD and hyperhomocysteinemia, but only S-AA decreased after vitamin supplementation (35). Homocysteine activates nuclear factor- in endothelial cells, possibly via oxidative stress (58), and increases monocyte chemoattractant protein-1 expression in vascular smooth muscle cells (59). Additionally, it stimulates interleukin-8 expression in human endothelial cultures (60). These inflammatory factors are known to participate in the development of atherosclerosis. Taken together, these reports suggest an association of elevated tHcy and low-grade inflammation in CAD. 

Carmody et al. found that the addition of homocysteine to a culture of vascular smooth muscle cells resulted in a dose-dependent increase in DNA synthesis and cell proliferation, but vitamins B6 and B 2 alone did not substantially inhibit the effect of homocysteine. However, the addition of folic acid resulted in significant inhibition of DNA synthesis (64). Rosiglitazone has been shown to reduce serum tHcy levels, smooth muscle proliferation, and intimal hyperplasia in Sprague-Dawley rats fed a diet high in methionine (65). 

Ebersole BJ, Diglio CA, Kaufman DW, Berg KA. 5-Hydroxytryptaminerlike receptors linked to increases in intracellular calcium concentration and inhibition of cyclic AMP accumulation in cultured vascular smooth muscle cells derived from bovine basilar artery. J Pharmacol Exp Ther 1993 266 692-699. 

Berk, B. C., Vekshtein, V., Gordon, H. M., et al. 1989. Angiotensin II-stimulated protein synthesis in cultured vascular smooth muscle cells. Hypertension 13 305-314. 

Eguchi, S., Matsumoto, T., Motley, E. D., et al. 1996. Identification of an essential signaling cascade for mitogen-activated protein kinase activation by angiotensin II in cultured rat vascular smooth muscle cells. Possible requirement of Gq-mediated p21ras activation coupled to a Ca2+/calmodulin-sensitive tyrosine kinase. J Biol Chem 271 14169-14175. 

Grant, S. L., Lassegue, B., Griendling, K. K., et al. 2000. Specific regulation of RGS2 messenger RNA by angiotensin II in cultured vascular smooth muscle cells. Mol Pharmacol 57 460-467. 

Ishizaka, N., Alexander, R. W., Laursen, J. B., et al. 1997. G protein-coupled receptor kinase 5 in cultured vascular smooth muscle cells and rat aorta. Regulation by angiotensin II and hypertension. J Biol Chem 272 32482-32488. 

Naftilan, A. J., Pratt, R. E., and Dzau, V. J. 1989. Induction of platelet-derived growth factor A-chain and c-myc gene expressions by angiotensin II in cultured rat vascular smooth muscle cells. J Clin Invest 83 1419-1424. 

Williams, B., P. Tsai, and R.W. Schrier. 1992. Glucose-induced downregulation of angiotensin II and arginine vasopressin receptors in cultured rat aortic vascular smooth muscle cells. Role of protein kinase C. J. Clin. Invest. 90 1992-1999. 

---