Gudjonsson T, Villadsen R, Rønnov-Jessen L, Petersen O W
Molecular and Cell Biology Research Laboratory, Icelandic Cancer Society, P.O. Box 5420, 125, Reykjavik, Iceland.
Cell Mol Life Sci. 2004 Oct;61(19-20):2523-34. doi: 10.1007/s00018-004-4167-z.
Defining the key players in normal breast differentiation is instrumental to understanding how morphogenesis becomes defective during breast cancer progression. During the past 2 decades much effort has been devoted to the development of technologies for purification and expansion of primary human breast cells in culture and optimizing a relevant microenvironment, which may help to define the niche that regulates breast differentiation and morphogenesis. In contrast to the general property of cancer, normal human cells have a finite lifespan. After a defined number of population doublings, normal cells enter an irreversible proliferation-arrested state referred to as replicative senescence. To overcome this obstacle for continuous long-term studies, replicative senescence can be bypassed by treatment of cells with chemical agents such as benzopyrene, by radiation or by transfection with viral oncogenes or the gene for human telomerase (human telomerase reverse transcriptase, hTERT). A drawback of some of these protocols is a concurrent introduction of chromosomal changes, which sometimes leads to a transformed phenotype and selection of a subpopulation, which may not be representative of the tissue of origin. In recent years, we have sought to establish immortalized primary breast cells, which retain crucial characteristics of their original in situ tissue pattern. This review discusses various approaches to immortalization of breast-derived epithelial and stromal cells and the application of such cell lines for studies on human breast morphogenesis.
明确正常乳腺分化过程中的关键参与者，有助于理解在乳腺癌进展过程中形态发生是如何出现缺陷的。在过去20年里，人们付出了巨大努力来开发用于纯化和扩增培养中的原代人乳腺细胞的技术，并优化相关的微环境，这可能有助于确定调节乳腺分化和形态发生的生态位。与癌症的一般特性不同，正常人类细胞的寿命是有限的。在经过一定数量的群体倍增后，正常细胞进入一种不可逆的增殖停滞状态，即复制性衰老。为了克服这一障碍以进行持续的长期研究，可以通过用化学试剂（如苯并芘）处理细胞、辐射或用病毒癌基因或人类端粒酶基因（人类端粒酶逆转录酶，hTERT）转染来绕过复制性衰老。这些方法中的一些缺点是会同时引入染色体变化，这有时会导致转化表型并选择一个亚群，而这个亚群可能不代表起源组织。近年来，我们一直在寻求建立永生化的原代乳腺细胞，这些细胞保留了其原始原位组织模式的关键特征。本综述讨论了乳腺来源的上皮细胞和基质细胞永生化的各种方法，以及此类细胞系在人类乳腺形态发生研究中的应用。
