Formation of Native-Type Collagen Fibrils by Cultured Fibroblasts.

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1. Fibroblasts were cultured for a long period of time, processed for electron microscopy, and the ultra-thin sections were examined under electron microscopes. Culture in the presence of long-lasting ascorbate (ascorbate phosphate) showed an increased mass of cells and extracellular matrices compared to the culture without the ascorbate. Collagen fibrils with periodic banding pattern were detected in the extracellular spaces, however, the banding images are not clear enough to identify the polarity of the fibrils or to asign the composition of constituting alpha chains.2. Collagen extracted from cultured cells formed segment-long-spacing crystallites (SLS) by dialysis against ATP, electron microscopic banding patterns of which were clear enough for further analyses.3. The SLS preparation made by collagen synthesized by cultured fibroblasts from an osteogenesis imperfecta patient showed normal as well as abnormal crystallites. Ultrastructue of the molecular aggregates suggested the site of irregularity.4. Fibrils were formed from the extracted collagens in vitro, showing the native-type banding pattern.5. Computer programs were developed to analyse, and simulate the banding patterns of collagen fibrils and SLS by the primary structure.