Characterization of rat alveolar type II cells in vitro by immunological, biochemical, and morphological criteria

James B. Mcmahon, Adaline C. Smith, Anthony Del Campo, Gurmukh Singh, Sikandar L. Katyal, Hildegard M. Schuller

Research output: Contribution to journalArticle

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Abstract

Using an anti-rat surfactant apoprotein antiserum which specifically reacts with cytoplasmic structures in alveolar type II cells on histopathology sections of rat lung, we have examined the immunoreactivity of pulmonary type II cells in vitro. Single cell suspensions of lung tissue were prepared from male Fischer 344 rats by intratracheal elastase digestion according to standard published methods. Cytocentrifuged preparations of the resulting cell suspensions revealed that approximately 40% of the cells stained positive for surfactant apoprotein using an immunoperoxidase staining technique. Without further cell fractionation steps, the cell suspensions were plated at colonial densities in growth medium. The cells that attached after 24 hours of incubation and at daily intervals were analyzed for surfactant apoprotein immunoreactivity as well as for proliferation, morphology, and phospholipid biosynthesis. The percentage of immunopositive cells increased with time from 75% at day 1 to 94% at 4 days after plating. This increase was paralleled by a linear increase in the number of immunopositive cells, which expanded into cell colonies. During the initial 5 days in vitro, the immunopositive cells retained their epithelial morphology and contained cytoplasmic osmiophilic bodies. Phospholipid biosynthesis by the isolated lung cells was analyzed and the data revealed that the rate of incorporation of 14C-choline into phosphatidylcholine increased with time in culture. These studies indicated that the anti-rat surfactant apoprotein antisera can be used to identify and quantitate functional alveolar type II cells in vitro. Thus the specific antisera may facilitate studies of type II cells undergoing various environmental alterations both in vivo and in vitro.

Original languageEnglish (US)
Pages (from-to)263-275
Number of pages13
JournalExperimental Lung Research
Volume11
Issue number4
DOIs
StatePublished - Jan 1 1986
Externally publishedYes

Fingerprint

Alveolar Epithelial Cells
Apoproteins
Surface-Active Agents
Rats
Immune Sera
Suspensions
Biosynthesis
Phospholipids
Pancreatic Elastase
Fractionation
Choline
Phosphatidylcholines
Plating
Lung
Tissue
In Vitro Techniques
Cytoplasmic Structures
Cell Fractionation
Inbred F344 Rats
Immunoenzyme Techniques

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry

Cite this

Characterization of rat alveolar type II cells in vitro by immunological, biochemical, and morphological criteria. / Mcmahon, James B.; Smith, Adaline C.; Campo, Anthony Del; Singh, Gurmukh; Katyal, Sikandar L.; Schuller, Hildegard M.

In: Experimental Lung Research, Vol. 11, No. 4, 01.01.1986, p. 263-275.

Research output: Contribution to journalArticle

Mcmahon, James B. ; Smith, Adaline C. ; Campo, Anthony Del ; Singh, Gurmukh ; Katyal, Sikandar L. ; Schuller, Hildegard M. / Characterization of rat alveolar type II cells in vitro by immunological, biochemical, and morphological criteria. In: Experimental Lung Research. 1986 ; Vol. 11, No. 4. pp. 263-275.
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