Formation of Native-Type Collagen Fibrils by Cultured Fibroblasts.

培养的成纤维细胞形成天然型胶原原纤维。

• 批准号: 01570008
• 负责人: KOBAYASHI Kunihiko
• 金额: $ 1.28万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570008/
• 关键词: Collagen; Fibroblasts; Electron microscope; Segment-long-spacing; Osteogensis imperfecta; Ascorbate; Computer simulation; Collagen fibrils; 細胞培養; コラ-ゲン分子束; アスコルビン酸2リン酸; 天然型コラ-ゲン

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.

1.将成纤维细胞长期培养，进行电子显微镜处理，并在电子显微镜下观察超薄切片。与不含抗坏血酸盐的培养物相比，在长效抗坏血酸盐（抗坏血酸磷酸盐）存在下的培养物显示出细胞和细胞外基质的质量增加。在细胞外间隙中检测到具有周期性带型的胶原原纤维，但带型图像不够清晰，无法确定原纤维的极性或组成α链的组成.从培养细胞中提取的胶原蛋白经ATP透析后形成节段长间距微晶（SLS），其电镜带型清晰，便于进一步分析.由成骨细胞合成的胶原蛋白制成的SLS制剂显示正常和异常的微晶。分子聚集体的超微结构显示了不规则的部位.胶原蛋白在体外形成原纤维，呈天然型带型.开发了计算机程序来分析和模拟胶原纤维和SLS的一级结构的带型。

项目成果

K. Kobayashi: "Direct visualization of affected collagen molecules synthesized by cultured fibroblasts from an osteogenesis imperfecta patient." Biochem. Biophys. Res. Commun.172(1). 217-222 (1990)

K. Kobayashi：“直接可视化由成骨不全症患者培养的成纤维细胞合成的受影响的胶原蛋白分子。”

KOBAYASHI,Kunihiko: "Direct visualization of affected collagen molecules synthesized by cultured fibroblasts from an osteogenesis imperfecta patient." Biochem.Biophys.Res.Commun.172. 217-222 (190)

KOBAYASHI，Kunihiko：“直接可视化由成骨不全患者培养的成纤维细胞合成的受影响的胶原蛋白分子。”

小林 邦彦: "点突然変異による異常コラ-ゲン分子の出現ーー電子顕微鏡による異常分子検出の試み [話題]" 医学のあゆみ. 155. 375 (1990)

Kunihiko Kobayashi：“由于点突变而出现异常胶原分子 - 尝试使用电子显微镜检测异常分子[主题]”医学史 155. 375 (1990)。

小林 邦彦: "点突然変異による異常コラ-ゲン分子の出現ーー電子顕微鏡による異常分子検出の試み〔話題〕" 医学のあゆみ. 155. 375 (1990)

Kunihiko Kobayashi：“点突变导致异常胶原分子的出现 - 尝试使用电子显微镜检测异常分子[主题]”医学史 155. 375 (1990)。

KOBAYASHI,Kunihiko: "Direct visualization of affected collagen molecules synthesized by cultured fibroblasts from an osteogenesis imperfecta patient." Biochem Biophys.Res.Commun.172. 217-222 (1990)

KOBAYASHI，Kunihiko：“直接可视化由成骨不全患者培养的成纤维细胞合成的受影响的胶原蛋白分子。”