Summary
Lung tissue expiants from mid-trimester human abortuses were maintained for 8 days in organ culture in medium with or without serum. Before the start of culture the cells lining the pre-alveolar ducts were undifferentiated and contained no lamellar bodies, the intracellular organelle that contains surfactant. After 4 days in organ culture, the epithelium lining the pre-alveolar ducts was composed of differentiated type II cells containing numerous lamellar bodies. During the 8-day culture period there was increased incorporation of [3H]choline into phosphatidylcholine and disaturated phosphatidylcholine. In addition, the specific activity of phosphatidate phosphohydrolase, a regulatory enzyme in lung phospholipid synthesis, increased 4-fold during the culture period. Lamellar bodies isolated by differential centrifugation from expiants maintained in culture for 7 days had the characteristic ultrastructure described for this organelle. Lamellar bodies were isolated from expiants which had been incubated with [14C]glycerol. When the glycerophospholipid composition of lamellar bodies was analyzed it was found that the majority of the radiolabeled glycerol (74%) was incorporated into phosphatidylcholine and into the anionic phospholipids, phosphatidylglycerol (5%) and phosphatidylinositol (6%). Thus, human fetal lung expiants maintained in organ culture contain differentiated type II cells which synthesize surfactant characteristic of human fetal lung at 36 to 38 weeks of gestation.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Adamson IYR, Bowden DH (1974) The type 2 cell as progenitor of alveolar epithelial regeneration: A cytodynamic study in mice after exposure to oxygen. Lab Invest 30:35–42
Bleasdale JE, Johnston JM (1981) Phosphatidic acid production and utilization. In: Farrell P (ed) The Developmental Biology of the Lung and the Pathobiology of Hyaline Membrane Disease. Academic Press, New York (In press)
Bligh EC, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917
Ekelund L, Arvidson G, Emanuelsson H, Myhrberg H, Astedt B (1975) Effect of cortisol on human fetal lung in organ culture. A biochemical, electron microscopic, and autoradiographic study. Cell Tissue Res 163:262–272
Farrell PM, Avery ME (1975) Hyaline membrane disease. Am Rev Resp Dis 111:657–688
Farrell PM, Hamosh M (1978) The biochemistry of fetal lung development. Clin. Perinatol. 5:197–229
Funkhouser JD, Hughes ER, Peterson RDA (1976) An organ culture system for study of fetal lung development. Biochem Biophys Res Commun 70:630–637
Gil J, Reiss OK (1973) Isolation and characterization of lamellar bodies and tubular myelin from rat lung homogenates. J Cell Biol 58:152–171
Hallman M, Kulovich M, Kirkpatrick E, Sugarman RG, Gluck L (1976) Phosphatidylinositol and phosphatidylglycerol in amniotic fluid: Indices of lung maturity. Am J Obstet Gynecol 125:613–617
Hallman M, Feldman BH, Kirkpatrick E, Gluck L (1977) Absence of phosphatidylglycerol (PG) in respiratory distress syndrome in the newborn. Study of the minor surfactant phospholipids in newborns. Pediat Res 11:714–720
Hallman M, Gluck L (1980) Formation of acidic phospholipids in rabbit lung during perinatal development. Pediatr Res 14:1250–1259
Hook GER, Gilmore LB, Tombropoulos EG, Fabro SE (1978) Fetal lung lamellar bodies in human amniotic fluid. Am Rev Respir Dis 117:541–550
Ikegami M, Silverman J, Adams FH (1979) Restoration of lung pressure-volume characteristics with various phospholipids. Pediatr Res 13:777–780
Johnston JM, Reynolds G, Wylie MB, MacDonald PC (1978) The phosphohydrolase activity in lamellar bodies and its relationship to phosphatidylglycerol and lung surfactant formation. Biochim Biophys Acta 531:65–71
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Mason RJ, Nellenbogen J, Clements JA (1976) Isolation of disaturated phosphatidylcholine with osmium tetroxide. J Lipid Res 17:281–284
Oulton M, Martin TR, Faulkner GT, Stinson D, Johnson JP (1980) Developmental study of a lamellar body fraction isolated from human amniotic fluid. Pediatr Res 14:722–728
Rousseau-Merck MF, Wyllie L, Basset F, Nezelof C (1976) In vitro behavior of human fetal lung maintained in organ culture. Virchows Arch (Pathol Anat) 371:305–321
Skipski VP, Barclay M (1969) Thin layer chromatography oflipids. In: Lowenstein JM (ed) Methods in Enzymology. Academic Press, N.Y., Vol. 14, pp. 530–598
Snyder JM, Mendelson CR, Johnston JM (1981) The effect of cortisol on rabbit fetal lung maturation in vitro. Devel Biol 85:129–140
Spitzer HL, Rice JM, MacDonald PC, Johnston JM (1975) Phospholipid biosynthesis in lung lamellar bodies. Biochem Biophys Res Comm 66:17–23
Stahlman MT, Gray ME (1978) Anatomical development and maturation of the lungs. Clin Perinatol 5:181–196
Stratton CJ (1978) The ultrastructure of multilamellar bodies and surfactant in the human lung. Cell Tissue Res 193:219–229
Yavin E, Zutra A (1977) Separation and analysis of 32P-labeled phospholipids by a simple and rapid thin-layer Chromatographic procedure and its application to cultured neuroblastoma cells. Anal Biochem 80:430–437
Author information
Authors and Affiliations
Additional information
This work was supported by USPH Grant 1 PO1 HD13912 and the Robert A. Welch Foundation, Houston, Texas
Rights and permissions
About this article
Cite this article
Snyder, J.M., Johnston, J.M. & Mendelson, C.R. Differentiation of type II cells of human fetal lung in vitro. Cell Tissue Res. 220, 17–25 (1981). https://doi.org/10.1007/BF00209962
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00209962