Freiberger H, Grove D, Sivarajah A, Pinnell S R
J Invest Dermatol. 1980 Nov;75(5):425-30. doi: 10.1111/1523-1747.ep12524080.
Human skin fibroblast culture promises to be a useful system for the investigation of the regulation of collagen biosynthesis and the study of abnormalities in collagen biosynthesis in connective tissue disorders. The effect of culture conditions on procollagen I biosynthesis has been determined. Optimal conditions for collagen biosynthesis were: 10% dialyzed heat-inactivated fetal calf serum, 0.15 mM ascorbate, and 0.078 mM beta-aminopropionitrile. Newly synthesized procollagen I accumulated in the medium at a linear rate for 18 hr. Preincubation of cells in labelling media for 4 hr before adding radioactive proline enhanced synthesis. Collagenase digestion was used to study overall collagen biosynthesis. 94% of all collagen synthesized was found in the medium, and 6% in the cell pellet. Under optimal conditions, collagen comprised 24% of all protein in the medium, and 14% of protein produced by the whole culture.
人类皮肤成纤维细胞培养有望成为一种有用的系统，用于研究胶原蛋白生物合成的调控以及结缔组织疾病中胶原蛋白生物合成异常的研究。已确定培养条件对前胶原I生物合成的影响。胶原蛋白生物合成的最佳条件为：10%经透析热灭活的胎牛血清、0.15 mM抗坏血酸盐和0.078 mMβ-氨基丙腈。新合成的前胶原I以线性速率在培养基中积累18小时。在添加放射性脯氨酸之前，将细胞在标记培养基中预孵育4小时可增强合成。使用胶原酶消化来研究整体胶原蛋白生物合成。发现所有合成的胶原蛋白中有94%存在于培养基中，6%存在于细胞沉淀中。在最佳条件下，胶原蛋白占培养基中所有蛋白质的24%，占整个培养物产生的蛋白质的14%。
