Ohki K, Nagayama A
J Cell Physiol. 1983 Mar;114(3):291-301. doi: 10.1002/jcp.1041140307.
Three cell lines from resident macrophages of BALB/c mice and four from activated macrophages of the same strain were isolated by infection with simian virus 40 (SV40). A majority of these cells showed dependency on L cell-conditioned medium (LCM), which is necessary for proliferation of normal macrophages in vitro. Somatic cell hybridization was applied in the study of macrophage growth responsiveness. A macrophage cell line (BR15) with strict dependency on LCM for growth was fused to a Chinese hamster cell line (hs222-16); it was found that dependency on LCM was a dominant trait in the hybrids. Following fusion of a macrophage cell line (BAM3) which grew without LCM to hs222-16, a large number of colonies appeared in the selection medium containing LCM. Four hybrids not requiring LCM for growth were selected in an LCM-free culture, and their hybrid properties were examined. Three out of the four hybrids secreted colony-stimulating factor (CSF) constitutively, whereas the fourth secreted no CSF. The level of acid phosphatase activity in the hybrids was higher than in the parent cells. Two peaks of CSF activity were observed after gel filtration chromatography of conditioned medium: One was eluted at molecular weight of 36,000 and the other at 17,000.
通过用猿猴病毒40(SV40)感染,从BALB/c小鼠的常驻巨噬细胞中分离出3种细胞系,从同一品系的活化巨噬细胞中分离出4种细胞系。这些细胞中的大多数表现出对L细胞条件培养基(LCM)的依赖性,LCM是体外正常巨噬细胞增殖所必需的。体细胞杂交被应用于巨噬细胞生长反应性的研究。将对LCM生长有严格依赖性的巨噬细胞系(BR15)与中国仓鼠细胞系(hs222-16)融合;发现对LCM的依赖性在杂种中是显性性状。将无需LCM即可生长的巨噬细胞系(BAM3)与hs222-16融合后,在含有LCM的选择培养基中出现了大量菌落。在无LCM培养中选择了4种无需LCM即可生长的杂种,并对它们的杂种特性进行了检测。4种杂种中有3种组成性分泌集落刺激因子(CSF),而第4种不分泌CSF。杂种中酸性磷酸酶活性水平高于亲本细胞。对条件培养基进行凝胶过滤层析后观察到两个CSF活性峰:一个在分子量36,000处洗脱,另一个在17,000处洗脱。