Biran S, Horowitz A T, Fuks Z, Vlodavsky I
Int J Cancer. 1983 May 15;31(5):557-66. doi: 10.1002/ijc.2910310506.
A routine procedure has been developed for the establishment in culture of normal primary and secondary human mammary epithelial cells. The high (80-100%) rate of success resulted from the combined use of a serum-free medium supplemented with high-density lipoprotein (HDL) and of cell plating on a naturally produced extracellular matrix (ECM). Plating on ECM greatly improved cell attachment, plating efficiency and initial outgrowth. HDL supported epithelial cell proliferation and prevented their detachment and degeneration while the omission of serum prevented the growth of stromal fibroblasts. Under these conditions we obtained from each specimen, and regardless of the patient's age, pure and actively dividing epithelial cell cultures forming a tightly packed and non-overlapping cell monolayer covering the entire area of the culture dish. These epithelial cultures could be easily dissociated and subcultured at a split ratio of 1:10. The described procedure will promote studies on the role of hormones and growth factors in the proliferation and differentiation of human mammary epithelial cells and on the susceptibility of human breast epithelial cells to various transforming agents and anti-cancer treatments.
已经开发出一种常规程序，用于将正常人乳腺原代和继代上皮细胞进行体外培养。高达80%-100%的成功率源于联合使用添加了高密度脂蛋白（HDL）的无血清培养基，以及将细胞接种在天然生成的细胞外基质（ECM）上。接种到ECM上极大地改善了细胞贴壁、接种效率和最初的生长。HDL支持上皮细胞增殖，并防止其脱离和退化，而不添加血清则可防止基质成纤维细胞生长。在这些条件下，无论患者年龄如何，我们从每个标本中都获得了纯净且活跃分裂的上皮细胞培养物，这些细胞形成紧密堆积且不重叠的细胞单层，覆盖培养皿的整个区域。这些上皮细胞培养物可以很容易地以1:10的传代比例解离并传代培养。所描述的程序将促进关于激素和生长因子在人乳腺上皮细胞增殖和分化中的作用，以及人乳腺上皮细胞对各种转化因子和抗癌治疗的敏感性的研究。
