Otsuka T, Ogo T, Nakano T, Niiro H, Kuga S, Satoh H, Furukawa Y, Zipori D, Niho Y
First Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Stem Cells. 1994 Jul;12(4):409-15. doi: 10.1002/stem.5530120408.
The expression of c-kit ligand and interleukin 6 (IL-6) genes in mouse bone marrow-derived stromal cell lines was examined using quantitative polymerase chain reaction (PCR) analysis based on the design of an internal DNA control. The stromal cells studied included the 14F1.1 endothelial-adipocytes that support long-term hemopoiesis and two additional cell lines (MBA-1, MBA-13) which do not have this function. All the cell lines expressed c-kit ligand gene constitutively, and this expression was not increased by lectins. On the other hand, the expression of the IL-6 gene was markedly induced in all the lines by lipopolysaccharide (LPS) and by phorbol 12-myristate 13 acetate (PMA). The constitutive expression of c-kit ligand in 14F1.1 cells was the lowest among the three cell lines studied and could be increased by stimulation with IL-4. Thus, we observed some quantitative differences among the cell lines in their expression of cytokine genes. However, the unique capacity of 14F1.1 cells to support in vitro hemopoiesis cannot thus far be explained solely on the basis of the ability of these cells to secrete cytokines which are not produced by other stromal cell lines. c-kit ligand may be necessary, but its presence alone is not sufficient for 14F1.1 cells to support prolonged hemopoiesis.
基于内部DNA对照的设计，使用定量聚合酶链反应（PCR）分析检测小鼠骨髓来源的基质细胞系中c-kit配体和白细胞介素6（IL-6）基因的表达。所研究的基质细胞包括支持长期造血的14F1.1内皮脂肪细胞以及另外两种不具备此功能的细胞系（MBA-1、MBA-13）。所有细胞系均组成性表达c-kit配体基因，且这种表达不会因凝集素而增加。另一方面，脂多糖（LPS）和佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）可显著诱导所有细胞系中IL-6基因的表达。在研究的三种细胞系中，14F1.1细胞中c-kit配体的组成性表达最低，且可通过IL-4刺激而增加。因此，我们观察到细胞系之间在细胞因子基因表达上存在一些定量差异。然而，迄今为止，14F1.1细胞支持体外造血的独特能力不能仅仅基于这些细胞分泌其他基质细胞系不产生的细胞因子的能力来解释。c-kit配体可能是必需的，但仅其存在不足以使14F1.1细胞支持长期造血。
