Chapter 5 Maintenance of Human and Rat Pulmonary Type II Cells in an Organotypic Culture System

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

“…Comparable morphological changes have been described for rat [9,10] and human [11] lung maintained in organotypic culture. This technique exploits the tendency of isolated cells from proteolytically dissociated fetal tissues to re-aggregate with other similar cell types in vitro [16,17].…”

“…As lamellar bodies are the sites of surfactant assembly and storage before secretion [3,4], and PC16 :0/16:0 is responsible for the surface-tension-lowering properties of surfactant [5], both these parameters have been extensively used for the assessment of tissue culture models of fetal lung development. Two such models are organ [6][7][8] and organotypic [9][10][11] cultures of immature fetal lung, for both of which changes have been described that resemble apparent developmental maturations in vitro. Results in the present paper compare for the first time analyses of morphology and phosphatidylcholine metabolism in parallel organ and organotypic cultures derived from human fetal lung.…”

“…One potential advantage of the organotypic cultures is the greater duration of cell viability in an apparently differentiated form [18]. Metabolic integrity and morphological structure were maintained for up to 4 weeks for human fetal lung in organotypic culture [11].…”

Lung Cell Biology

“…This deterioration is a difficulty with differentiated cells from adults. In their thorough review of type I1 cells in vitro, Douglas et al (59) describe ways of overcoming this limitation so as to have long-term cultures available. They derived from an adult rat a clonal cell line L-2 that retains type I1 cell characteristics for a certain time but loses them after -35 population doublings in vitro.…”

“…Fetal lung cells are dissociated enzymatically; under suitable organotypic culture conditions they reaggregate to form histotypic elements resembling epithelial tubes in the fetal lung (58,59). In such cultures alveolarlike structures are formed that are lined by cells resembling type I1 cells and by attenuated cells similar to type I cells (58,59). These cultures perform the metabolic functions of surfactant biosynthesis and secrete the product into the internal alveolar spaces where it is transformed to tubular myelin.…”