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-smooth muscle actin expression
by primary human lung fibroblasts
1 Lung Cell Kinetics Laboratory and The
Cardiovascular Institute, Michael Reese Hospital, Chicago, Illinois
60616; and 3 Facultad de Ciencias,
Primary human lung fibroblasts were separated
into small ( group I),
intermediate ( group II), and large
( group III) subpopulations by unit
gravity sedimentation (1 G). The three subsets retained differences in
cell size for up to 15 days of primary culture. Flow cytometric
(fluorescence-activated cell sorter) measurements of forward-angle
light scatter agreed well with fibroblast volume measured by image
analysis and confirmed the utility of forward-angle light scatter for
discriminating size subpopulations. Group
II fibroblasts accumulated most rapidly by 8 days of
culture and also contained the greatest proportion of S and
G2/M phase cells as determined by
fluorescence-activated cell sorter. Fibroblasts that were
immunoreactive with antibodies to 
-smooth muscle actin (-SMA)
were found only in group III. In situ
end labeling of fragmented DNA detected apoptotic cells in both
groups II and III, but double labeling for in situ
end labeling and -SMA revealed apoptotic cells in both the
-SMA-positive and -negative populations. These results demonstrate
that primary human lung fibroblasts behave as predicted by classic
models of cell cycle progression and differentiation. However, they do
not support the hypothesis that the expression of -actin is related
to apoptosis. We also describe a simple and reproducible method for the
high-yield isolation of human lung fibroblast subsets of differing
proliferative potential and phenotype.
lung cells; myofibroblast; apoptosis; proliferation; cell heterogeneity
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