Fujii M, Yoshino I, Suzuki M, Higuchi T, Mukai S, Aoki T, Fukunaga T, Sugimoto Y, Inoue Y, Kusuda J, Saheki T, Sato M, Hayashi S, Tamaki M, Sugano T
Department of Biochemical Science and Technology, Faculty of Agriculture, Kagoshima University, Japan.
Int J Biochem Cell Biol. 1996 Dec;28(12):1381-91. doi: 10.1016/s1357-2725(96)00076-3.
To study liver functions of chicken, we examined the primary culture of chicken hepatocytes, and found an easy method of long-term culture with free atmosphere exchange. Chicken hepatocytes were obtained by collagenase perfusion and cultured at 37 degrees C as a monolayer without substratum in serum-free L-15 medium (pH 7.8) with free atmosphere exchange. The amounts of albumin and transferrin in medium were assayed by ELISA. The culture of chicken hepatocytes was maintained in the serum-free L15-medium )pH 7.) and 37 degrees C with free atmosphere exchange for 20 days. The amount of albumin secreted in the medium decreased to low levels early in culture; however, this was followed by marked increase from day 9 to day 17 of culture. The amount of transferrin was constant until day 6, then it too increased with further culture. We reported an easy method for the simple monolayer culture of chicken hepatocytes in serum-free L12 medium (pH 7.8) with free atmosphere exchange over an extended period. Expression of liver-specific functions, viz. albumin and transferrin synthesis, was observed after 1 week of culture.
为研究鸡的肝功能,我们检测了鸡肝细胞的原代培养,发现了一种可进行自由气体交换的长期培养的简便方法。通过胶原酶灌注获取鸡肝细胞,并在37℃下于无血清L-15培养基(pH 7.8)中不使用基质进行单层培养,且可进行自由气体交换。采用酶联免疫吸附测定法(ELISA)检测培养基中白蛋白和转铁蛋白的含量。鸡肝细胞在无血清L15培养基(pH 7)中于37℃下进行自由气体交换培养20天。培养基中分泌的白蛋白量在培养早期降至低水平;然而,随后在培养的第9天至第17天显著增加。转铁蛋白量在第6天前保持恒定,之后随着进一步培养也增加。我们报道了一种在无血清L12培养基(pH 7.8)中进行鸡肝细胞简单单层培养的简便方法,可进行自由气体交换并持续较长时间。培养1周后观察到肝脏特异性功能即白蛋白和转铁蛋白合成的表达。