AJP - Cell Physiology, Vol 254, Issue 2 C235-C242, Copyright © 1988 by American Physiological Society

ARTICLES

Effect of seeding density and time in culture on vascular smooth muscle cell proteins

C. L. Seidel, V. White, C. Wallace, J. Amann, D. Dennison, L. A. Schildmeyer, B. Vu, J. C. Allen, L. Navarro and S. Eskin
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030.

The purpose of this work was to determine the effect of seeding density and time in culture on quantitative and qualitative characteristics of myosin in primary cultures of vascular smooth muscle cells (VSMCs). Enzymatically dispersed VSMCs from femoral arteries and saphenous veins of adult dogs were seeded at a density of 10(3)-10(5) cells/cm2 and assayed after 7 days or at 10(5) cells/cm2 and assayed between 1 and 10 days. Myosin, actin, and total protein contents as well as electrophoretic and immunoreactive characteristics of myosin heavy chains (MHCs) were determined. Total and contractile protein contents were independent of seeding density and increased with time in culture. Freshly dispersed cells exhibited two MHCs (MHC-1 and MHC-2) but, within 24 h after culturing, only cells attached to the dish expressed a third protein band (MHC-3), which had electrophoretic mobility and immunoreactivity similar to purified platelet MHC. MHC-3 appeared before onset of cell division and, by 4 days in culture when cells were proliferating, became the dominant MHC form. Loss of MHC-1 and MHC-2 could be prevented by growing cells in a serum-free, defined media that prevented proliferation. These data indicate that seeding density does not affect myosin content, but that with time in culture expression of a MHC with characteristics similar to nonmuscle myosin occurs. Expression of MHC-3 is associated with cell attachment, whereas loss of MHC-1 and MHC-2 requires proliferation.

This article has been cited by other articles:

				A. M. Kahn, A. Husid, T. Odebunmi, J. C. Allen, C. L. Seidel, and T. Song Insulin inhibits vascular smooth muscle contraction at a site distal to intracellular Ca2+ concentration Am J Physiol Endocrinol Metab, May 1, 1998; 274(5): E885 - E892. [Abstract] [Full Text] [PDF]

				A. M. Kahn, A. Husid, J. C. Allen, C. L. Seidel, and T. Song Insulin Acutely Inhibits Cultured Vascular Smooth Muscle Cell Contraction by a Nitric Oxide Synthase–Dependent Pathway Hypertension, October 1, 1997; 30(4): 928 - 933. [Abstract] [Full Text]