L-PROLINE IS AN ESSENTIAL AMINO-ACID FOR HEPATOCYTE GROWTH IN CULTURE
L-PROLINE IS AN ESSENTIAL AMINO-ACID FOR HEPATOCYTE GROWTH IN CULTURE
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DOI：
10.1016/0006-291x(84)91173-2
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发表时间：
1984-01-01
影响因子：
3.1
通讯作者：
ICHIHARA, A
中科院分区：
文献类型：
--
作者：
NAKAMURA, T;TERAMOTO, H;ICHIHARA, A
为改善胶原包被培养皿原代培养成年大鼠肝细胞的条件，研究了不同商品培养基对胰岛素加表皮生长因子刺激的细胞DNA复制合成的影响。脯氨酸缺乏的培养基，如Leibovitz's L-15，Spille's最小必需培养基和Dulbecco's改良最小必需培养基，不能诱导肝细胞的DNA合成，而富含脯氨酸的培养基，如威廉姆斯培养基E，McCoy's 5A和Ham's F-12，能显著诱导肝细胞增殖。当脯氨酸缺乏的培养基中补充L-脯氨酸时，细胞合成DNA响应这两种激素。顺式-4-羟基-L-脯氨酸强烈抑制DNA合成的诱导，而不影响细胞的蛋白质合成或显示任何细胞毒性。通过向培养基中加入过量的脯氨酸，这种抑制被完全恢复。添加培养基中不存在的其他氨基酸并不促进DNA合成。L-脯氨酸通过影响细胞内胶原的合成，对诱导培养的肝细胞增殖显然是必需的。
For improvement of the culture conditions of adult rat hepatocytes in primary culture in collagen coated dishes, effects of various commercial culture media on the induction of replicative DNA synthesis of the cells stimulated by insulin plus epidermal growth factor were studied. Proline-deficient media, such as Leibovitz''s L-15, Eagle''s minimal essential medium and Dulbecco''s modified minimal essential medium, did not induce DNA synthesis in hepatocytes, whereas proline-rich media, such as Williams medium E, McCoy''s 5A and Ham''s F-12, induced markedly hepatocyte proliferation. When the proline-deficient media were supplement with L-proline, the cells synthesized DNA in response to the 2 hormones. Cis-4-hydroxyl-L-proline strongly inhibited the induction of DNA synthesis, without affecting protein synthesis of the cells or showing any cytotoxicity. This inhibition was recovered completely by adding excess proline to the medium. Addition of other amino acids not present in the medium did not promote DNA synthesis. L-proline apparently is essential for induction of hepatocyte proliferation in culture, through its affect on synthesis of intracellular collagen.