Maintaining intact mature lung tissue in culture using low melt agarose

Autor: P. Marlene Absher, John N. Evans, Frank Casty, Paul Shapiro
Rok vydání: 1995
Předmět:
Zdroj: Methods in Cell Science. 17:245-249
ISSN: 1573-0603
1381-5741
DOI: 10.1007/bf00986229
Popis: The development of methods to culture intact viable tissue is integral to understanding mechanisms of disease. Mature lung tissue, in particular, is difficult to maintain for any extended period of time in culture due to collapse of airspaces and rapid tissue necrosis. In the present stud, we report the methodology for culturing whole lung tissue in vitro using low-melt agarose to maintain lung airspaces open and viable. Perfused rat lungs were inflated in situ with a warm liquid low-melt agarose solution. Upon cooling, the agarose polymerizes maintaining the airspaces in a open state and allows thin sections of lung to be cut and cultured in a defined environment. Viability of the cultured lung tissue was assessed biochemically. Protein synthesis was maintained for 7 days in cultured lung tissue and was dependent upon the oxygen concentration in which the tissue was cultured. Cell proliferation was assessed immunohistochemically in parenchyma of lung slices cultured in the presence of 5-bromo-2′-deoxyuridine (BRDU). BRDU labeling indices for tissue cultured in 21% O2 or greater showed an oxygen-dependent increase in cell proliferation after 3 days in culture followed by a return to baseline levels after 7 days. The data describe the maintenance of mature lung tissue in culture and indicate the usefulness of the in vitro lung slice model for examining the mechanistic basis of lung injury and subsequent remodeling.
Databáze: OpenAIRE
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