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岩藻糖和氨基葡萄糖掺入细胞结合及培养基释放的大分子中。

Incorporation of fucose and glucosamine into cell bound and medium released macromolecules.

作者信息

Herrmann H, Havaranis A S, Doetschman T C

出版信息

J Cell Physiol. 1975 Jun;85(3):557-68. doi: 10.1002/jcp.1040850307.

Abstract

(1) Determinations were carried out on the incorporation of fucose-6-(3H) and glucosamine-6-(3H) into trichloracetic acid insoluble macromolecules which remained bound to the cells or were released into the medium of chick embryo muscle cell cultures. The radioactivity determined in the medium was corrected for unspecific binding of label to components of the medium. (2) During an incorporation period of six hours the incorporation per microgram DNA with fucose as label into cell bound macromolecules is about twice as high as the incorporation into macromolecules released into medium. With glucosamine about twice as much is incorporated into medium released into the cell bound macromolecules. (3) The incorporation per microgram DNA increased during a culture period of three days but the increase ceases at different times during this culture period when determined with fucose or glucosamine or for cell bound and medium released material. (4) An increase in cell density increases the incorporation per DNA of fucose and to a much slighter extent that of glucosamine. Reduction of cell density by addition of cytosine arabinoside to the medium does not increase the incorporation per microgram DNA. (5) The effect of changes of fibroblast/myoblast ratios on the incorporation of fucose and glucosamine were examined. No significant effect was observed for a ratio of 10-30% fibroblasts when control cultures or cultures after cell sedimentation were maintained in complete medium. Marked changes were observed after culture in medium without protein components. Under these conditions an increase in the fibroblast/myoblast ratios were observed as well as an increase in the incorporation of label into medium released and a decrease into cell bound macromolecules.

摘要

(1) 对岩藻糖 -6-(³H) 和葡糖胺 -6-(³H) 掺入三氯乙酸不溶性大分子的情况进行了测定,这些大分子要么仍与鸡胚肌细胞培养物中的细胞结合,要么释放到培养基中。对培养基中测定的放射性进行了校正,以消除标记物与培养基成分的非特异性结合。(2) 在六小时的掺入期内,以岩藻糖为标记物时,每微克 DNA 掺入细胞结合大分子中的量约为掺入释放到培养基中的大分子的两倍。以葡糖胺为标记物时,掺入释放到培养基中的量约为掺入细胞结合大分子的两倍。(3) 在三天的培养期内,每微克 DNA 的掺入量增加,但在用岩藻糖或葡糖胺测定时,或者对于细胞结合和释放到培养基中的物质,在这个培养期的不同时间增加停止。(4) 细胞密度的增加会增加每 DNA 中岩藻糖的掺入量,而葡糖胺的掺入量增加幅度要小得多。向培养基中添加阿糖胞苷降低细胞密度并不会增加每微克 DNA 的掺入量。(5) 研究了成纤维细胞/成肌细胞比例变化对岩藻糖和葡糖胺掺入的影响。当对照培养物或细胞沉淀后的培养物在完全培养基中维持时,对于 10 - 30% 的成纤维细胞比例未观察到显著影响。在无蛋白质成分的培养基中培养后观察到明显变化。在这些条件下,观察到成纤维细胞/成肌细胞比例增加,同时释放到培养基中的标记物掺入量增加,而细胞结合大分子中的掺入量减少。

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