Stackpole C W, Cremona P, Kassel R L
J Immunol Methods. 1979;30(3):231-43. doi: 10.1016/0022-1759(79)90097-8.
A one-step isopycnic density gradient centrifugation procedure is described for purification of highly viable and homogeneous tumor cells from a variety of solid mouse tumors. Mechanically suspended cells are layered onto preformed continuous gradients of medium 199-buffered 7--33% metrizamide (density range 1.05--1.20 g/cu. cm) isoosmotic with mouse plasma and centrifuged for 30 min. Large numbers of tumor cells, generally 85--95% viable and free from 80--95% of contaminating host lymphoid and phagocytic cells and erythrocytes, were consistently recovered from fractionated thymomas, melanomas, and fibrosarcomas. By a variety of criteria, cell surface and other biological properties of gradient-purified tumor cells were normal.
本文描述了一种一步等密度梯度离心法，用于从多种小鼠实体瘤中纯化高活性且均一的肿瘤细胞。将机械悬浮的细胞铺在预先形成的连续梯度的199培养基缓冲的7%-33%的甲泛葡胺（密度范围为1.05-1.20 g/cm³）上，该梯度与小鼠血浆等渗，然后离心30分钟。从分馏的胸腺瘤、黑色素瘤和纤维肉瘤中持续回收了大量肿瘤细胞，这些细胞通常85%-95%具有活性，且80%-95%不含宿主淋巴细胞、吞噬细胞和红细胞等污染物。通过多种标准评估，梯度纯化的肿瘤细胞的细胞表面和其他生物学特性均正常。
