Shikita M, Tsuneoka K, Hagiwara S, Tsurufuji S
J Cell Physiol. 1981 Oct;109(1):161-9. doi: 10.1002/jcp.1041090118.
Seven days after subcutaneous injection of 2% carrageenin solution in mice, the induced inflammatory tissue was isolated and treated with 0.1% trypsin. When the dispersed cells were incubated in a nutrient medium containing 5-20% calf serum, the cells grew adhering to the culture-dish surface and colony-stimulating factor (CSF) was accumulated in the medium gradually. Addition of lipopolysaccharide (Escherichia coli endotoxin) in the cell culture did not enhance the CSF production. It was shown by isoelectrofocusing that the majority of the produced CSF was an acidic molecule (pI = 3.8), while the treatment of this CSF with neuraminidase yielded a less acidic CSF species (pI = 5.1). Upon gel-filtration chromatography in the presence of 6 M guanidine HCl, the CSF exhibited an apparent molecular weight of 42,000. On the other hand, polyacrylamide gel-electrophoresis in the presence of 0.1% sodium dodecyl sulfate gave a molecular weight estimate of 65,000. Microscopic examination of the bone marrow cell culture showed that about one-third of the colonies were granulocytic. Addition of prostaglandin E1(PGE1) in the bone marrow cell culture significantly inhibited the action of the CSF, while prostaglandin D2 was less inhibitory than PGE1. The result suggests that the cells isolated from the inflammatory tissue are capable of producing CSF, which may have a role for proliferation and maturation of the mononuclear phagocytes at the site of inflammation.
给小鼠皮下注射2%角叉菜胶溶液7天后，分离诱导产生的炎症组织并用0.1%胰蛋白酶处理。当将分散的细胞在含有5%-20%小牛血清的营养培养基中培养时，细胞贴附于培养皿表面生长，并且集落刺激因子（CSF）逐渐在培养基中积累。在细胞培养中添加脂多糖（大肠杆菌内毒素）并未增强CSF的产生。等电聚焦显示，所产生的CSF大多数是酸性分子（pI = 3.8），而用神经氨酸酶处理该CSF可产生酸性较弱的CSF种类（pI = 5.1）。在6 M盐酸胍存在下进行凝胶过滤层析时，CSF的表观分子量为42,000。另一方面，在0.1%十二烷基硫酸钠存在下进行聚丙烯酰胺凝胶电泳，估计分子量为65,000。对骨髓细胞培养物进行显微镜检查显示，约三分之一的集落为粒细胞集落。在骨髓细胞培养中添加前列腺素E1（PGE1）可显著抑制CSF的作用，而前列腺素D2的抑制作用比PGE1弱。结果表明，从炎症组织分离的细胞能够产生CSF，这可能对炎症部位单核吞噬细胞的增殖和成熟起作用。
