Serum-free cultures of murine adrenal cortical cells
Serum-free cultures of murine adrenal cortical cells
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DOI：
10.1016/j.jpedsurg.2006.08.027
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发表时间：
2006-12-01
影响因子：
2.4
通讯作者：
Dunn, James C. Y.
中科院分区：
文献类型：
--
作者：
Chu, Yinting;Wu, Benjamin M.;Dunn, James C. Y.
目的：探讨小鼠肾上腺皮质细胞移植前培养的可行性；方法：原代小鼠肾上腺皮质细胞保存在含有胎牛和马血清的培养基或无血清培养基中。采用实时聚合酶链式反应定量培养细胞中肾上腺皮质基因表达水平。结果：使用含血清培养基导致培养物中许多细胞生长，但此类培养物中Sf-1、Dax-1和Cyp11b1的表达迅速下降。相反，在无血清培养基中没有明显的细胞生长。在无血清培养基中培养小鼠肾上腺皮质细胞导致 Sf-1、Dax-1 和 Cyp11b1 基因表达水平较高。在无血清培养基中，肾上腺皮质细胞也以剂量依赖性方式通过增加Cyp11b1的表达并抑制Dax-1的表达来响应促肾上腺皮质激素。在无血清培养基中添加碱性成纤维细胞生长因子可使Sf-1、Dax-1和Cyp11b1的表达维持4周。结论：从成年小鼠分离的肾上腺皮质细胞在含碱性成纤维细胞生长因子的无血清培养基中成功维持。该培养系统可能适合在移植前在体外进一步操作肾上腺皮质细胞。 (c) 2006 Elsevier Inc. 保留所有权利。
Purpose: The feasibility of culturing murine adrenal cortical cells before transplantation was Adrenal cortical cells; investigated in this study.Method: Primary murine adrenal cortical cells were maintained in either fetal bovine and horse sera containing media or serum-free media. Real-time polymerase chain reaction was used to quantify the levels of adrenal cortical gene expression in the cultured cells.Results: The use of sera-containing media led to the growth of many cells in the culture, but the expression of Sf-1, Dax-1, and Cyp11b1 in such cultures declined rapidly. In contrast, there was no significant cell growth in the serum-free culture medium. Culturing murine adrenal cortical cells in the serum-free medium resulted in higher levels of Sf-1, Dax-1, and Cyp11b1 gene expression. In the serum-free medium, adrenal cortical cells also responded to adrenocorticotropic hormone by increasing the expression of Cyp11b1 and suppressing the expression of Dax-1 in a dose-dependent manner. The addition of basic fibroblast growth factor to the serum-free medium maintained the expression of Sf-1, Dax-1, and Cyp11b1 for 4 weeks.Conclusion: Adrenal cortical cells isolated from adult mice were successfully maintained in a serum-free culture medium with basic fibroblast growth factor. This culture system may be suitable for further manipulation of adrenal cortical cells in vitro before transplantation. (c) 2006 Elsevier Inc. All rights reserved.