Danielson K G, Knepper J E, Kittrell F S, Butel J S, Medina D, Durban E M
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
In Vitro Cell Dev Biol. 1989 Jun;25(6):535-43. doi: 10.1007/BF02623566.
Clonal populations were isolated from the mouse mammary cell line, COMMA-D, by transfection with a dominant-selectable gene, pSV2Neo, which confers resistance to the antibiotic, G418. Seven of twenty-four clones isolated retained the ability of the parental line to repopulate cleared mammary fat pads in vivo as ductal-alveolar hyperplasias. Two sublines designated CDNR2 and CDNR4 retained hyperplastic growth potential after multiple passages in vitro with low incidence of tumor formation. A third subpopulation, CDNR1, contained a single integration site for the pSV2Neo plasmid indicating a bonafide clonal origin for this subline. CDNR1 cells displayed heterogeneous growth phenotypes in vivo including hyperplasia, adenocarcinoma, and bone formation. Functional differentiation of CDNR1 cells organized as alveolarlike structures in vivo or on floating collagen gels in vitro was observed as determined by immunoperoxidase staining for the milk-specific protein, casein. Overall, the results indicate that a subset of cells from the COMMA-D cell line may be functionally analogous to stem cells existing in the mammary gland.
通过用显性选择基因pSV2Neo转染赋予对抗生素G418抗性的小鼠乳腺细胞系COMMA-D，分离出克隆群体。分离出的24个克隆中有7个保留了亲代细胞系在体内重新填充清除的乳腺脂肪垫形成导管-腺泡增生的能力。两个指定为CDNR2和CDNR4的亚系在体外多次传代后仍保留增生生长潜力，肿瘤形成发生率较低。第三个亚群CDNR1含有pSV2Neo质粒的单个整合位点，表明该亚系有真正的克隆起源。CDNR1细胞在体内表现出异质性生长表型，包括增生、腺癌和骨形成。通过对乳特异性蛋白酪蛋白进行免疫过氧化物酶染色确定，观察到在体内或体外漂浮胶原凝胶上组织成肺泡样结构的CDNR1细胞的功能分化。总体而言，结果表明COMMA-D细胞系中的一部分细胞可能在功能上类似于乳腺中存在的干细胞。
