Kimball P C, Boehm-Truitt M, Schochetman G, Schlom J
J Natl Cancer Inst. 1976 Jan;56(1):111-7. doi: 10.1093/jnci/56.1.111.
Primary cell cultures of mammary tumors from Rill, GR, DD, BALB/cfC3H, and BALB/c mice were prepared by trypsin-EDTA dissociation of tumors. Cultures from these strains contained predominantly cells of epithelial morphology which formed three-dimensional domelike structures. Cultures from Rill, GR, DD, and BALB/cfC3H tumors produced extra-cellular type-B mouse mammary tumor virus(es) (MuMTV), either in the absence of detectable type-C virus or with less than 1% contamination with type-C virus. This was determined by radioimmunoassays for MuMTV and murine leukemia virus (MuLV) antigens. Only BALB/c cultures produced MuMTV with as much as 3% contaminating MuLV. High levels of MuMTV surface antigen were also found in soluble form in culture supernatants. Virus polypeptide analyses by electrophoresis on polyacrylamide gels showed that the Rill BALB/cfC3H, DD, and BALB/c viruses all contained polypeptides characteristic of MuMTV. Primary cultures of mammary tumor cells make available a source of purified MuMTV antigens, structural proteins, and nucleic acids for comparative studies of MuMTV from various mouse strains.
通过用胰蛋白酶 - 乙二胺四乙酸(EDTA)解离肿瘤,制备了来自里尔(Rill)、GR、DD、BALB/cfC3H和BALB/c小鼠的乳腺肿瘤原代细胞培养物。这些品系的培养物主要包含上皮形态的细胞,这些细胞形成三维穹顶状结构。来自里尔、GR、DD和BALB/cfC3H肿瘤的培养物产生细胞外B型小鼠乳腺肿瘤病毒(MuMTV),要么在没有可检测到的C型病毒的情况下,要么C型病毒污染少于1%。这是通过对MuMTV和鼠白血病病毒(MuLV)抗原的放射免疫测定来确定的。只有BALB/c培养物产生的MuMTV有高达3%的MuLV污染。在培养上清液中也发现了高水平的可溶性MuMTV表面抗原。通过聚丙烯酰胺凝胶电泳进行的病毒多肽分析表明,里尔、BALB/cfC3H、DD和BALB/c病毒都含有MuMTV特有的多肽。乳腺肿瘤细胞的原代培养物为来自各种小鼠品系的MuMTV的比较研究提供了纯化的MuMTV抗原、结构蛋白和核酸的来源。