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. 1988 Jul 15;253(2):451–457. doi: 10.1042/bj2530451

A comparison of the specificity of phosphatidylcholine synthesis by human fetal lung maintained in either organ or organotypic culture.

P A Caesar 1, S J Wilson 1, C S Normand 1, A D Postle 1
PMCID: PMC1149319  PMID: 3178723

Abstract

Human fetal lung (14-18 weeks gestation) was maintained in either organ or organotypic culture. By 4 days in organ culture or 14 days in organotypic culture, epithelial cells within both culture systems exhibited well-developed apical microvilli and possessed numerous intracellular lamellar bodies characteristic of surfactant phospholipid stores. However, analysis of the pattern of synthesis of individual molecular species of phosphatidylcholine by [14C]choline incorporation and reversed-phase h.p.l.c. showed that this apparent maturation was not paralleled by an increased synthesis of the dipalmitoyl species in either culture system. By contrast, the fractional synthesis of dipalmitoyl phosphatidylcholine, expressed as a percentage of total [14C]choline incorporation, decreased with time in both organ and organotypic culture. Moreover, these fractions were not significantly different from those measured in parallel monolayer cultures of mixed human fetal lung cells that displayed mainly fibroblast morphology. These results suggest that the synthesis pattern of phosphatidylcholine species by lung cells in culture is determined principally by their incubation conditions and not by their state of apparent maturation.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adams F. H., Fujiwara T., Emmanouilides G. C., Räihä N. Lung phospholipids of human fetuses and infants with and without hyaline membrane disease. J Pediatr. 1970 Nov;77(5):833–841. doi: 10.1016/s0022-3476(70)80244-x. [DOI] [PubMed] [Google Scholar]
  2. Aeberhard E. E., Barrett C. T., Kaplan S. A., Scott M. L. Stimulation of phosphatidylcholine synthesis by fatty acids in fetal rabbit type II pneumocytes. Biochim Biophys Acta. 1986 Jan 3;875(1):6–11. doi: 10.1016/0005-2760(86)90004-4. [DOI] [PubMed] [Google Scholar]
  3. Arvidson G. A. Fractionation of naturally occurring lecithins according to degree of unsaturation by thin-layer chromatography. J Lipid Res. 1965 Oct;6(4):574–577. [PubMed] [Google Scholar]
  4. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  5. Ballard P. L., Ertsey R., Gonzales L. K., Liley H. G., Williams M. C. Isolation and characterization of differentiated alveolar type II cells from fetal human lung. Biochim Biophys Acta. 1986 Sep 4;883(2):335–344. doi: 10.1016/0304-4165(86)90326-0. [DOI] [PubMed] [Google Scholar]
  6. Bangham A. D. Lung surfactant: how it does and does not work. Lung. 1987;165(1):17–25. doi: 10.1007/BF02714417. [DOI] [PubMed] [Google Scholar]
  7. Batenburg J. J., Funkhouser J. D., Klazinga W., Van Golde L. M. On the suitability of organotypic cultures of fetal rat lung type II cells for biochemical studies concerning development. Biochim Biophys Acta. 1983 Jan 7;750(1):60–67. doi: 10.1016/0005-2760(83)90204-7. [DOI] [PubMed] [Google Scholar]
  8. Batenburg J. J. Isolated type II cells from fetal lung as model in studies on the synthesis and secretion of pulmonary surfactant. Lung. 1980;158(4):177–192. doi: 10.1007/BF02713722. [DOI] [PubMed] [Google Scholar]
  9. Batenburg J. J., Longmore W. J., van Golde L. M. The synthesis of phosphatidylcholine by adult rat lung alveolar type II epithelial cells in primary culture. Biochim Biophys Acta. 1978 Apr 28;529(1):160–170. doi: 10.1016/0005-2760(78)90114-5. [DOI] [PubMed] [Google Scholar]
  10. Batenburg J. J., den Breejen J. N., Yost R. W., Haagsman H. P., van Golde L. M. Glycerol 3-phosphate acylation in microsomes of type II cells isolated from adult rat lung. Biochim Biophys Acta. 1986 Oct 3;878(3):301–309. doi: 10.1016/0005-2760(86)90237-7. [DOI] [PubMed] [Google Scholar]
  11. Blank M. L., Spector A. A., Kaduce T. L., Snyder F. Composition and incorporation of [3H]arachidonic acid into molecular species of phospholipid classes by cultured human endothelial cells. Biochim Biophys Acta. 1986 Jun 11;877(1):211–215. doi: 10.1016/0005-2760(86)90136-0. [DOI] [PubMed] [Google Scholar]
  12. Brumley G. W., Chernick V., Hodson W. A., Normand C., Fenner A., Avery M. E. Correlations of mechanical stability, morphology, pulmonary surfactant, and phospholipid content in the developing lamb lung. J Clin Invest. 1967 May;46(5):863–873. doi: 10.1172/JCI105586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chevalier G., Collet A. J. In vivo incorporation of choline- 3 H, leucine- 3 H and galactose- 3 H in alveolar type II pneumocytes in relation to surfactant synthesis. A quantitative radoautographic study in mouse by electron microscopy. Anat Rec. 1972 Nov;174(3):289–310. doi: 10.1002/ar.1091740303. [DOI] [PubMed] [Google Scholar]
  14. Douglas W. H., Moorman G. W., Teel R. W. The formation of histotypic structures from monodisperse fetal rat lung cells cultured on a three-dimensional substrate. In Vitro. 1976 May;12(5):373–381. doi: 10.1007/BF02796315. [DOI] [PubMed] [Google Scholar]
  15. Douglas W. H., Sanders R. L., Hitchcock K. R. Maintenance of human and rat pulmonary type II cells in an organotypic culture system. Methods Cell Biol. 1980;21A:79–94. doi: 10.1016/s0091-679x(08)60759-1. [DOI] [PubMed] [Google Scholar]
  16. Ekelund L., Arvidson G., Emanuelsson H., Myhrberg H., Astedt B. Effect of cortisol on human fetal lung in organ culture: a biochemical, electron-microscopic and autoradiographic study. Cell Tissue Res. 1975 Nov 12;163(3):263–272. doi: 10.1007/BF00219463. [DOI] [PubMed] [Google Scholar]
  17. Engle M. J., Sanders R. L., Douglas W. H. Type II alveolar cells in organotypic culture. A model system for the study of surfactant synthesis. Biochim Biophys Acta. 1980 Feb 22;617(2):225–236. doi: 10.1016/0005-2760(80)90165-4. [DOI] [PubMed] [Google Scholar]
  18. GROVER J. W. The influence of age and environmental factors on the behaviour of reaggregated embryonic lung cells in culture. Exp Cell Res. 1962 Mar;26:344–359. doi: 10.1016/0014-4827(62)90187-8. [DOI] [PubMed] [Google Scholar]
  19. Gonzales L. W., Ballard P. L., Ertsey R., Williams M. C. Glucocorticoids and thyroid hormones stimulate biochemical and morphological differentiation of human fetal lung in organ culture. J Clin Endocrinol Metab. 1986 Apr;62(4):678–691. doi: 10.1210/jcem-62-4-678. [DOI] [PubMed] [Google Scholar]
  20. Gross I., Smith G. J., Maniscalco W. M., Czajka M. R., Wilson C. M., Rooney S. A. An organ culture model for study of biochemical development of fetal rat lung. J Appl Physiol Respir Environ Exerc Physiol. 1978 Sep;45(3):355–362. doi: 10.1152/jappl.1978.45.3.355. [DOI] [PubMed] [Google Scholar]
  21. Hayashi Y., Urade R., Kito M. Distribution of phospholipid molecular species containing arachidonic acid and cholesterol in V79-UF cells. Biochim Biophys Acta. 1987 Apr 24;918(3):267–273. [PubMed] [Google Scholar]
  22. King R. J., Gikas E. G., Ruch J., Clements J. A. The radioimmunoassay of pulmonary surface active material in sheep lung. Am Rev Respir Dis. 1974 Sep;110(3):273–281. doi: 10.1164/arrd.1974.110.3.273. [DOI] [PubMed] [Google Scholar]
  23. Longmuir K. J., Bleasdale J. E., Quirk J. G., Johnston J. M. Regulation of lamellar body acidic glycerophospholipid biosynthesis in fetal rabbit lung in organ culture. Biochim Biophys Acta. 1982 Aug 18;712(2):356–364. doi: 10.1016/0005-2760(82)90353-8. [DOI] [PubMed] [Google Scholar]
  24. Mason R. J., Nellenbogen J., Clements J. A. Isolation of disaturated phosphatidylcholine with osmium tetroxide. J Lipid Res. 1976 May;17(3):281–284. [PubMed] [Google Scholar]
  25. Meyrick B., Reid L. Electron microscopic aspects of surfactant secretion. Proc R Soc Med. 1973 Apr;66(4):386–387. [PMC free article] [PubMed] [Google Scholar]
  26. Patterson C. E., Davis K. S., Beckman D. E., Rhoades R. A. Fatty acid synthesis in the fetal lung: relationship to surfactant lipids. Biochim Biophys Acta. 1986 Aug 14;878(1):110–126. doi: 10.1016/0005-2760(86)90349-8. [DOI] [PubMed] [Google Scholar]
  27. Patton G. M., Fasulo J. M., Robins S. J. Separation of phospholipids and individual molecular species of phospholipids by high-performance liquid chromatography. J Lipid Res. 1982 Jan;23(1):190–196. [PubMed] [Google Scholar]
  28. Postle A. D. Method for the sensitive analysis of individual molecular species of phosphatidylcholine by high-performance liquid chromatography using post-column fluorescence detection. J Chromatogr. 1987 Apr 10;415(2):241–251. doi: 10.1016/s0378-4347(00)83216-8. [DOI] [PubMed] [Google Scholar]
  29. Rüstow B., Kunze D., Rabe H., Reichmann G. The molecular species of phosphatidic acid, diacylglycerol and phosphatidylcholine synthesized from sn-glycerol 3-phosphate in rat lung microsomes. Biochim Biophys Acta. 1985 Jul 31;835(3):465–476. doi: 10.1016/0005-2760(85)90116-x. [DOI] [PubMed] [Google Scholar]
  30. Scott J. E., Possmayer F., Quirie M. A., Tanswell A. K., Harding P. G. Alveolar pre-type II cells from the fetal rabbit lung: effect of confluence on the production of disaturated phosphatidylcholine. Biochim Biophys Acta. 1986 Dec 5;879(3):292–300. doi: 10.1016/0005-2760(86)90218-3. [DOI] [PubMed] [Google Scholar]
  31. Snyder J. M., Johnston J. M., Mendelson C. R. Differentiation of type II cells of human fetal lung in vitro. Cell Tissue Res. 1981;220(1):17–25. doi: 10.1007/BF00209962. [DOI] [PubMed] [Google Scholar]
  32. Spector A. A., Kiser R. E., Denning G. M., Koh S. W., DeBault L. E. Modification of the fatty acid composition of cultured human fibroblasts. J Lipid Res. 1979 May;20(4):536–547. [PubMed] [Google Scholar]
  33. Spector A. A., Yorek M. A. Membrane lipid composition and cellular function. J Lipid Res. 1985 Sep;26(9):1015–1035. [PubMed] [Google Scholar]
  34. Sterzel W., Bedford P., Eisenbrand G. Automated determination of DNA using the fluorochrome Hoechst 33258. Anal Biochem. 1985 Jun;147(2):462–467. doi: 10.1016/0003-2697(85)90299-4. [DOI] [PubMed] [Google Scholar]
  35. Young S. L., Tierney D. F. Dipalmitoyl lecithin secretion and metabolism by the rat lung. Am J Physiol. 1972 Jun;222(6):1539–1544. doi: 10.1152/ajplegacy.1972.222.6.1539. [DOI] [PubMed] [Google Scholar]

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