Characterization of the biophysical properties of human tumor and bone marrow cells as a preliminary step to the use of centrifugal elutriation in autologous bone marrow transplantation

Abstract

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 distribution 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 metastic to BM.