AJP - Cell Physiology, Vol 254, Issue 2 C338-C343, Copyright © 1988 by American Physiological Society
High-pressure pulsation of central and microvessel pulmonary artery endothelial cells
M. Rabinovitch, T. Bothwell, M. Mullen and B. N. Hayakawa
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
We developed an in vitro method of pulsating central and microvessel
pulmonary artery endothelial cells that would allow us to study the effects
of increased distending pressures over a prolonged period of time.
Preservation of the contact-inhibited monolayer was assessed on phase
contrast microscopy and, in addition, scanning and transmission electron
microscopy (SEM, TEM) were used to determine whether there were alterations
in the surface characteristics or intracytoplasmic organelles that
suggested cellular damage. The cells used were obtained from Rambouillet
lambs, age 3-5 days, anesthetized with halothane and ventilated. The
endothelium was harvested from the central pulmonary artery (CPA) by
scraping the luminal surface and from the microvessels (MPA) by infusing
microcarrier beads 40-140 microns external diameter. After the second
passage in culture, the cells were seeded onto the translucent, flexible
polyvinylchloride membrane of a transducer dome and grown to confluence.
The cell dome was then connected to a blank dome with an attached quartz
transducer, to a reservoir, and to stainless steel bellows tubing, all
filled with culture medium and affixed to a pulsation generator. By varying
the height of the reservoir, the amplitude of excursion of the bellows
tubing, and the rate, the cells could be pulsated at a given distending
pressure and frequency. Confluent CPA endothelial cells from three lambs
and MPA cells from two others were studied after pulsation at both 100/60
and 20/10 mmHg, 60 times/min for 48 h and after nonpulsation. On phase
contrast light microscopy and on SEM, the cells remained
confluent.(ABSTRACT TRUNCATED AT 250 WORDS)
