Elsevier

Experimental Hematology

Volume 38, Issue 12, December 2010, Pages 1219-1230
Experimental Hematology

Hematological Malignancy
Chronic lymphoid leukemia cells are highly sensitive to the combination of prednisolone and daunorubicin, but much less to doxorubicin or epirubicin

https://doi.org/10.1016/j.exphem.2010.09.001Get rights and content

Objective

To generate a comprehensive map of the drug sensitivity of chronic lymphoid leukemia cells (CLL) using a newly developed in vitro drug-sensitivity assay based on automated evaluation of cell viability on single-cell level.

Materials and Methods

Primary CLL cells from 77 patients were tested using automated digital fluorescence microscopy. The effect of 27 frequently used chemotherapeutic agents was measured in short-term fluorescence survival assay. To avoid typical in vitro artifacts such as growth factor depletion and oxidative damage, the cell were cultured in a novel, total human blood lysate−based medium (OmniSanguine) in order to preserve the composition of growth factor flora and redox conditions of the in vivo environment.

Results

CLL cells from different patients showed considerable heterogeneity in their drug-sensitivity patterns. This pattern was stable even after in vitro activation of cell proliferation. Half of the samples were sensitive to fludarabine and chlorambucil. Daunorubicin was the most potent drug. It was effective in 75 of 77 cases. In addition, daunorubicin and prednisolone showed a strong synergistic effect.

Conclusions

We suggest that the combination of low-dose daunorubicin and prednisolone might be an additional treatment option for therapy-resistant cases of CLL.

Section snippets

Isolation of CLL cells from peripheral blood

Five milliliters of total blood from CLL patients were drawn in heparinized glass tube for routine laboratory evaluation of blood chemistry and cellularity. All samples selected for this study contained high peripheral lymphoid cell counts (>20 × 103/μL). The superfluous material, usually between 1 and 2 mL, was shipped from the clinical centers to the Karolinska Laboratory by overnight mail at room temperature. The blood was diluted with equal volumes of phosphate-buffered saline and tumor

A new in vitro drug-sensitivity assay

CLL cells rapidly undergo apoptosis when cultured in vitro in conventional cell culture medium, which contrasts with their prolonged survival in vivo [26]. In this respect, CLL cells are similar to normal peripheral blood B cells. In vitro survival of B cells is mainly determined by the availability of the proper mixture of cytokines. Peripheral blood B cells and CLL B cells produce and secrete similar amounts of cytokines and cytokine receptors in vitro [27]. Depletion of cytokine-mediated

Discussion

Chronic lymphoid leukemia cells are favorite targets of in vitro chemosensitivity and chemoresistance assays due to easy availability in large quantities in the peripheral blood of CLL patients. These assays, however, are complicated by the limited survival of the lymphoma cells outside of the body, as well as by difficulty comparing in vitro and in vivo response rates. In the present article, we used a new in vitro cell culture system that maintains the composition of whole human blood but at

Acknowledgments

This work was supported by the Swedish Research Council, the Swedish Cancer Fund and Karolinska Institute Strategic Fund.

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