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Papers In Press, published online ahead of print June 1, 2006
J. Lipid Res., doi:10.1194/jlr.M600054-JLR200

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Journal of Lipid Research, Vol. 47, 1322-1331, June 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Lipidomics of cellular and secreted phospholipids from differentiated human fetal type II alveolar epithelial cells

Anthony D. Postle^1,*, Linda W. Gonzales, Wolfgang Bernhard, Graeme T. Clark^*, Marye H. Godinez^**, Rodolfo I. Godinez^** and Philip L. Ballard

^* Division of Infection, Inflammation, and Repair, School of Medicine, University of Southampton, Southampton, UK
Department of Pediatrics, Division of Neonatology, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA
Department of Neonatology, Children's Hospital, Medical Faculty, Eberhard-Karls-University, Tübingen, Germany
^** Department of Anesthesiology and Critical Care Medicine, Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA

The online version of this article (available at http://www.jlr.org) contains additional six tables.

Published, JLR Papers in Press, March 2, 2006.

¹ To whom correspondence should be addressed. e-mail: adp{at}soton.ac.uk

Maturation of fetal alveolar type II epithelial cells in utero is characterized by specific changes to lung surfactant phospholipids. Here, we quantified the effects of hormonal differentiation in vitro on the molecular specificity of cellular and secreted phospholipids from human fetal type II epithelial cells using electrospray ionization mass spectrometry. Differentiation, assessed by morphology and changes in gene expression, was accompanied by restricted and specific modifications to cell phospholipids, principally enrichments of shorter chain species of phosphatidylcholine (PC) and phosphatidylinositol, that were not observed in fetal lung fibroblasts. Treatment of differentiated epithelial cells with secretagogues stimulated the secretion of functional surfactant-containing surfactant proteins B and C (SP-B and SP-C). Secreted material was further enriched in this same set of phospholipid species but was characterized by increased contents of short-chain monounsaturated and disaturated species other than dipalmitoyl PC (PC16:0/16:0), principally palmitoylmyristoyl PC (PC16:0/14:0) and palmitoylpalmitoleoyl PC (PC16:0/16:1). Mixtures of these PC molecular species, phosphatidylglycerol, and SP-B and SP-C were functionally active and rapidly generated low surface tension on compression in a pulsating bubble surfactometer. These results suggest that hormonally differentiated human fetal type II cells do not select the molecular composition of surfactant phospholipid on the basis of saturation but, more likely, on the basis of acyl chain length.

Supplementary key words phosphatidylcholine • phosphatidylinositol • electrospray ionization mass spectrometry • differentiation

Abbreviations: DCI, dexamethasone + 8-bromo-cAMP + isobutylmethylxanthine; ESI-MS, electrospray ionization mass spectrometry; PA, phosphatidic acid; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PI, phosphatidylinositol; PS, phosphatidylserine; SP-A, -B or -C, surfactant proteins A, B or C

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