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expression with alveolar epithelial cell
phenotype in vitro
Will Rogers Institute Pulmonary Research Center, Division of Pulmonary and Critical Care Medicine, University of Southern California, Los Angeles, California 90033; and The Pulmonary Center, Boston University School of Medicine, Boston, Massachusetts 02118
T1
is a
recently identified gene expressed in the adult rat lung by alveolar
type I (AT1) epithelial cells but not by alveolar type II (AT2)
epithelial cells. We evaluated the effects of modulating alveolar
epithelial cell (AEC) phenotype in vitro on T1 expression using
either soluble factors or changes in cell shape to influence phenotype.
For studies on the effects of soluble factors on T1 expression, rat
AT2 cells were grown on polycarbonate filters in serum-free medium
(MDSF) or in MDSF supplemented with either bovine serum (BS, 10%), rat
serum (RS, 5%), or keratinocyte growth factor (KGF, 10 ng/ml) from
either day 0 or day
4 through day 8 in
culture. For studies on the effects of cell shape on T1 expression,
AT2 cells were plated on thick collagen gels in MDSF supplemented with
BS. Gels were detached on either day 1 (DG1) or day 4 (DG4) or were left
attached until day 8. RNA and protein were harvested at intervals between days
1 and 8 in culture,
and T1 expression was quantified by Northern and Western blotting, respectively. Expression of T1 progressively increases in AEC grown
in MDSF ± BS between day 1 and
day 8 in culture, consistent with
transition toward an AT1 cell phenotype. Exposure to RS or KGF from
day 0 prevents the increase in T1
expression on day 8, whereas addition
of either factor from day 4 through
day 8 reverses the increase. AEC
cultured on attached gels express high levels of T1 on
days 4 and
8. T1 expression is markedly
inhibited in both DG1 and DG4 cultures, consistent with both inhibition and reversal of the transition toward the AT1 cell phenotype. These
results demonstrate that both soluble factors and alterations in cell
shape modulate T1 expression in parallel with AEC phenotype and
provide further support for the concept that transdifferentiation between AT2 and AT1 cell phenotypes is at least partially reversible.
collagen gel; keratinocyte growth factor; rat serum; transdifferentiation; type I cell
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