Maturation of undifferentiated lung epithelial cells into type II cells in vitro: a temporal process that parallels cell differentiation in vivo.
BACKGROUND
Formation of alveolar-like structures (ALS) by mature fetal rabbit type II pneumocytes (day 29 gestation) and long-term differentiation on Engelbreth-Holms-Swarm mouse tumor extract or EHS gel (Matrigel) were reported by our group (Blau et al., 1988. J. Cell Physiol., 136:203-214). We now describe structural organization and differentiation of immature lung epithelial cells, isolated at day 22 gestation, into mature type II cells in vitro.


METHODS
Peripheral pulmonary tissue was pooled and undifferentiated epithelial cells isolated for primary culture on Matrigel. Cells were examined 12-16 h after plating and on days 1, 3, 5, and 7 of culture and assessed by phase contrast and by transmission electron microscopy after fixation in situ.


RESULTS
Cells formed ALS 12-16 h after plating. Spherule diameter increased about four to eight times from day 1-7 in culture. There was rapid transformation of tall columnar cells to cuboidal, normal polarization of cells with respect to cell-free lumen of ALS, progressive reduction of glycogen zones, apparent gradual increase of cell organelles such as Golgi apparatus, rough endoplasmic reticulum and mitochondria, and apparent extrusion of lipidic figures into the lumen. These morphologic transformations in vitro temporally paralleled cell differentiation in vivo. The relative increase of 14C-acetate precursor into phosphatidylcholine in contrast to cardiolipin was consistent with these transformations.


CONCLUSIONS
Under the conditions of our culture system, maturation of undifferentiated pulmonary epithelial cells is reproduced in vitro along the same time course and according to the same developmental sequence of fetal lungs in vivo.
Chinoy
M R
MR
Department of Cellular and Molecular Physiology, Hershey Medical Center, Hershey, Pennsylvania.
Antonio-Santiago
M T
MT
Scarpelli
E M
EM
D000085:Acetates / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D002855:Chromatography, Thin Layer; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005865:Gestational Age; D008168:Lung / Q000166:cytology* / Q000196:embryology / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D015388:Organelles / Q000648:ultrastructure; D010743:Phospholipids / Q000096:biosynthesis; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000196:embryology / Q000378:metabolism / Q000648:ultrastructure; D011817:Rabbits
D000085:Acetates; D010743:Phospholipids