Enzymatically dissociated fetal rat lung cells were frozen in medium containing dimethyl sulfoxide (DMSO) as the cryoprotectant, and stored at liquid nitrogen temperature. The cells were retrieved from frozen storage and seeded into Gelfoam collagen sponges. The cultures were examined morphologically using tetrazolium chloride (TTC) vital staining, light microscopy, and transmission electron microscopy, at various points throughout a two-week in vitro period. Fetal rat lung organotypic cultures established from previously frozen cells possessed the characteristics of cultures derived from freshly dissociated lung. Though the percent recovery of viable cells following freezing was low (<50%), those cells that survived the cryopreservation procedure exhibited histotypic reaggregation. These pulmonary epithelial cells formed alveolar-like structures (ALS) in which the cells associated to enclose a central lumen. The majority of these epithelial cells differentiated in vitro, developing the morphologic characteristics of type II alveolar epithelial cells. It is suggested that cryopreservation methods are applicable to in vitro studies on the pulmonary surfactant system.
|Original language||English (US)|
|Number of pages||11|
|Journal||Birth Defects: Original Article Series|
|State||Published - Dec 1 1980|
ASJC Scopus subject areas
- Developmental Biology