Culture of type II pneumocytes on a type II cell-derived fibronectin-rich matrix

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Abstract
Recent evidence suggests that during primary culture, type II pneumocytes synthesize and deposit components of an extracellular matrix. The present study investigated the response of freshly isolated type II cells to a preformed, fibronectin-rich matrix synthesized by type II cells over a 6-day interval of primary culture on a plastic surface. Type II cells on 6-day matrix (M6) degraded the preformed matrix and deposited newly synthesized fibronectin more rapidly than cells on plastic, suggesting that M6 itself stimulated type II cell-mediated matrix turnover. In type II cells on plastic, incorporation of radiolabeled thymidine into DNA increased 620 and 1,880% after 2 and 3 days in culture, respectively, as the cells assumed a more flattened phenotype. Although cells on M6 did not divide, both basal rates of thymidine labeling and sensitivity to serum modulators of DNA synthesis were enhanced by the M6 surface, as compared with plastic. Culture of type II cells on surfaces of purified fibronectin enhanced the rate of DNA synthesis in a manner similar to that observed on M6; this effect was blocked by antifibronectin. The data suggest that more rapid fibronectin synthesis and deposition are important components of the response of type II cells to primary culture. Extracellular matrix produced by type II cells appears to be similar to the basement membrane onto which these cells proliferate in vivo after lung injury. A fibronectin-rich surface in itself may thus induce additional extracellular matrix synthesis and further direct cellular differentiation and proliferation.