Keng P C, Rubin P, Constine L S, Frantz C, Nakissa N, Gregory P
Int J Radiat Oncol Biol Phys. 1984 Oct;10(10):1913-22. doi: 10.1016/0360-3016(84)90272-4.
The principle of centrifugal elutriation (CE) depends on a balance of an outwardly directed centrifugal force and inwardly directed fluid flow and buoyant forces. This method (CE) can be used effectively to separate cells on the basis of size. In the murine model, neoplastic cells from different tumors are generally larger than bone marrow cells and can be removed from bone marrow almost completely with centrifugal elutriation. In order to determine if CE is capable of eliminating human tumor cells from harvested bone marrow (BM), the biophysical characteristics of a variety of human tumor cells and bone marrow cells were determined. Human tumor cells were dispersed into single cell suspensions by several enzymatic digestion and mechanical dissociation methods. The size and density characteristics of these cells were determined with an electronic particle counter and channelyzer and density gradients. Of 40 solid tumors studied, 29 tumors had cell size distributions distinctively larger than BM, as was found in the experimental animal model. The cell size distributions of tumor cells from 11 solid tumors and 7 leukemias were not substantially different from that of BM. Mixtures of BM and cultured human hypernephroma, ovarian, and neuroblastoma cells, were separated into BM and tumor fractions by CE. The separation results as indicated by the labeling index and colony forming efficiency of tumor cells in each fraction showed that a BM fraction virtually free of tumor cells could be obtained. Thus, CE should be able to separate BM cells from most tumor cells metastatic to BM.
离心淘析(CE)的原理取决于向外的离心力与向内的流体流动及浮力之间的平衡。这种方法(CE)可有效地根据细胞大小分离细胞。在小鼠模型中,来自不同肿瘤的肿瘤细胞通常比骨髓细胞大,通过离心淘析几乎可以将其从骨髓中完全去除。为了确定CE是否能够从采集的骨髓(BM)中清除人类肿瘤细胞,我们测定了多种人类肿瘤细胞和骨髓细胞的生物物理特性。通过几种酶消化和机械解离方法将人类肿瘤细胞分散成单细胞悬液。用电子粒子计数器、通道分析仪和密度梯度测定这些细胞的大小和密度特征。在所研究的40种实体瘤中,有29种肿瘤的细胞大小分布明显大于骨髓,这与实验动物模型中的情况相同。11种实体瘤和7种白血病的肿瘤细胞的细胞大小分布与骨髓的细胞大小分布没有显著差异。通过CE将骨髓与培养的人类肾上腺肿瘤、卵巢肿瘤和神经母细胞瘤细胞的混合物分离成骨髓部分和肿瘤部分。各部分肿瘤细胞的标记指数和集落形成效率表明的分离结果显示,可以获得几乎不含肿瘤细胞的骨髓部分。因此,CE应该能够将骨髓细胞与大多数转移至骨髓的肿瘤细胞分离。