Investigating the relationship between cell cycle stage and diosgenin-induced megakaryocytic differentiation of HEL cells using sedimentation field-flow fractionation.

Differentiation therapy could be one strategy for stopping cancer cell proliferation. A plant steroid, diosgenin, is known to induce megakaryocytic differentiation in human erythroleukemia (HEL) cells. In recent studies, the use of sedimentation field-flow fractionation (SdFFF) allowed the preparation of subpopulations that may differ in regard to sensitivity to differentiation induction. The specific goal of this study was to determine the relationship between cell cycle stage and sensitivity to megakaryocytic differentiation induction of HEL cells. After first confirming the capacity of diosgenin to specifically select targets, hyperlayer SdFFF cell sorting was used to prepare fractions according to cell cycle position from crude HEL cells. The sensitivities of these fractions to diosgenin-induced differentiation were then tested. The coupling of SdFFF cell separation to imaging flow cytometry showed that G1-phase cells were more sensitive to differentiation induction than S/G2M-phase cells, confirming the relationship between cell status at the start of induction, the extent of the biological event, and the potential of SdFFF in cancer research.

Diosgenin dose-dependent apoptosis and differentiation induction in human erythroleukemia cell line and sedimentation field-flow fractionation monitoring.

To limit or stop cancer spreading, one of the most prevalent strategies is to induce cancer cell death. Differentiation therapy and apoptosis induction are two ways to achieve this goal. Sedimentation field-flow fractionation (SdFFF) has been described as an effective tool for cell separation, respecting integrity and viability. Because SdFFF takes advantage of intrinsic properties of eluted cells (size, density, shape), we studied the capacity of SdFFF to monitor specific biophysical modifications that occurred during cellular apoptosis or differentiation induction. Then, we used, as an in vitro cellular model of apoptosis and differentiation, diosgenin dose-dependent induction in the polyvalent human erythroleukemia cell line. Two other chemicals were used: phorbol myristate acetate (differentiation inducer) and staurosporine (apoptosis inducer). Our results demonstrated a correlation between SdFFF elution profile changes and induction of effective biological processes. Thus, after acquisition of a reference profile, SdFFF could be used alone to follow chemically induced biological events, suggesting many different applications such as testing series of molecules, evaluation of new cellular/biological models used in different life science fields, or sorting purified populations with the aim of better understanding mechanisms of induced cellular events.