Expression of liver specific functions in chicken hepatocytes

鸡肝细胞肝脏特异性功能的表达

• 批准号: 09660322
• 负责人: FUJII Makoto
• 金额: $ 2.18万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660322/
• 关键词: Chicken; Hepatocyte; Primary culture; Serum-free culture; Liver specific function; Depoisoning enzymes; Environmental pollution chemicals; アルブミン; トランスフェリン; 単層培養

Rats primary hepatocytes have been usually cultured in a 5% COィイD22ィエD2 incubator at pH 7.4. However, we found that chicken hepatocytes were able to cultured in serum-free L-15 medium for a long period in free gas exchange with the atmosphere (without COィイD22ィエD2 gas). The amount of albumin secreted in the medium decreased to low level early, followed by marked increase from day 9 to day 17 of culture. The profile of appearance of albumin m-RNA was very similar to that of albumin protein. Such observation was not observed in rat hepatocytes, showing some difference in the properties of hepatocytes between avian and rat. Therefore, we tried to monitored the effects of enbironmental pollution chemicals on metabolism in liver of wild birds using chicken hepatocytes as a model. 10ィイD1-1ィエD1 M TCDD (tetracholoro-dibenzo-p-dioxin) decreased the expression of albumin, and 10ィイD1-2ィエD1 M TCDD markedly induced the expression of the depoisoning enzyme, P450 CYP1A4.

大鼠原代肝细胞通常在pH 7.4的5% CO2-D22-D2培养箱中培养。然而，我们发现鸡肝细胞能够在无血清L-15培养基中与大气进行自由气体交换（没有CO_（22）CO_（22）D_2气体）的情况下长时间培养。白蛋白分泌量在培养早期下降到低水平，然后在培养的第9天至第17天显著增加。白蛋白m-RNA的外观轮廓与白蛋白蛋白的外观轮廓非常相似。在大鼠肝细胞中未观察到这种观察结果，表明禽类和大鼠之间的肝细胞性质存在一些差异。因此，我们尝试以鸡肝细胞为模型，监测环境污染物对野生鸟类肝脏代谢的影响。10 μ mol/L D1-1 μ mol/L D1 M TCDD（tetracoloro-dibenzo-p-dioxin，四氯二苯并二恶英）可降低白蛋白的表达，10 μ mol/L D1-2 μ mol/L D1 M TCDD可显著诱导解毒酶P450 CYP 1A 4的表达。

项目成果

Fujii et al.: "An economical method for primary culture of chicken hepatocytes in serum-free medium"Jpn. Poult. Sci.. 33. 15-22 (1996)

Fujii等人：“一种在无血清培养基中原代培养鸡肝细胞的经济方法”Jpn。

Fujii et al.,: "Primary culture of chicken hepatocytes in serum-free medium(pH7.8) secreted albumin and transferrin for a long period in free gas exchange with the atmosphere."Int,J.Biochem.Cell Biol.. 28. 1381-1391 (1996)

Fujii 等人：“在无血清培养基（pH7.8）中对鸡肝细胞进行原代培养，在与大气进行游离气体交换的过程中长时间分泌白蛋白和转铁蛋白。”Int,J.Biochem.Cell Biol.. 28

Inoue Y., Ose T., Mukai S., Ehira S., Yonekura M., Hou D-X., Fujii M.: "Inhibition of the acid lipase activity by apolipoprotein A-I in the presence of lysosomal proteases."Biosci. Biotechnol. Biochem.. 63. 937-939 (1999)

Inoue Y.、Ose T.、Mukai S.、Ehira S.、Yonekura M.、Hou D-X.、Fujii M.：“在溶酶体蛋白酶存在下，载脂蛋白 A-I 对酸性脂肪酶活性的抑制。”Biosci。

Fujii et al.,: "Primary culture of chicken hepatocytes in serum-free medium(pH7.8)secreted albumin and transferrin for a long period in free gas exchange with the atmosphere." Int.J.Biochem.Cell Biol.28. 1381-1391 (1966)

Fujii等人：“鸡肝细胞在无血清培养基（pH7.8）中进行原代培养，长时间与大气进行游离气体交换，分泌白蛋白和转铁蛋白。”

Fujii M., Yoshino I.., Suzuki M., Higuchi T., Mukai S. ,Aoki T. , Fukunaga T. , Sugimoto Y. , Inoue Y. , Kusuda J. , Saeki T . , Sato M. , Hayashi S. ,Tamaki M. , Sugano T.: "Primary culture of chicken hepatocytes in serum-free medium(pH 7.8) secreted alb

Fujii M.、Yoshino I..、Suzuki M.、Higuchi T.、Mukai S.、Aoki T.、Fukunaga T.、Sugimoto Y.、Inoue Y.、Kusuda J.、Saeki T.