Bronchial explant culture using an serum free medium was carried out on polystyrene dishes coated with different extracellular substrates. The NHBEC outgrowth area was then measured, and the effects of the extracellular substrates were compared. Experiment 1 : four types of collagen ; type I, type II, type III, and type IV, were compared. The outgrowth area (cm
2) of type I, type II, type III, and type IV was 24, 17, 25, and 28, respectively, in donor 1, with type IV>type II (P<0.05); 12, 11, 14, and 16, respectively, in donor 2, with type IV>type I-type II (P<0.05); 26, 19, 19, and 28, respectively, in donor 3, with no significant differences : 24, 17, 17, and 28, respectively, in donor 4, with type IV>type II-type III (P<0.05); and 16, 17, 15, and 23, respectively, in donor 5, with type IV>type I-type II-type III. Experiment 2 : observations were focused on type IV which was the most effective type in Experiment 1. Three groups were prepared by coating type IV with laminin (LM) [group L], fibronectin (Fn) [group F], and LM and FN [group LF], and then compared with Group C using only type IV. The outgrowth area (cm
2) of C, L, F, and LF, respectively was 13, 16, 17, and 20 with LF> C (P <0.05) in donor 6 ; 20, 23, 20, and 32, with LF> F-C (P<0.05) in donor 7 ; 16, 22, 23, and 28, with LF>C (P< 0.05) in donor 8 ; 19, 20, 19, and 29, with LF> L-F-C (P<0.05) in donor 9 ; and 17, 32, 23, and 28, with LF> F-C (P<0.05) in donor 10. The above-mentioned results showed that coating with LM, FN added to type IV had an effective action on the proliferation of NHBEC, suggested that NHBEC may be a useful substrate for coating the lumen of the artificial trachea.
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