Sedimentation field flow fractionation to study human erythroleukemia cell megakaryocytic differentiation after short period diosgenin induction

• Published in: Journal of Chromatography A Vol. 1157; no. 1; pp. 309 - 320
• Main Authors: Léger, D.Y., Battu, S., Liagre, B., Cardot, P.J.P., Beneytout, J.L.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 20.07.2007; Elsevier
• Subjects: Base Sequence; Biological and medical sciences; Cell Differentiation; Cell Line, Tumor; Cell separation; Chromatography, High Pressure Liquid; Differentiation; Diosgenin; Diosgenin - metabolism; DNA Primers; Fractionation, Field Flow; General aspects; Human erythroleukemia cells; Humans; Leukemia, Erythroblastic, Acute - pathology; Medical sciences; Megakaryocyte; Megakaryocytes - pathology; Ploidies; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm - isolation & purification; Sedimentation field flow fractionation; Spectrophotometry, Ultraviolet; Tumors; Washout; Sedimentation field flow fractionation; Washout; Diosgenin; Human erythroleukemia cells; Megakaryocyte; Cell separation; Differentiation; Erythroleukemia; Human; Steroid; Field flow fractionation; Cell differentiation; Separation method; Cell line; Spirostane derivatives; Kinetics; Tumor cell
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2007.04.051

Anti-cancer differentiation therapy could be one strategy to stop cancer cell proliferation. We propose a new sedimentation field flow fractionation (SdFFF) cell separation application in the field of cancer research. It concerns the study of megakaryocytic differentiation processes after a short exposure to an inducting agent (diosgenin). Washout process and early dual SdFFF separation – removing the influence of diosgenin and decreasing the influence of undifferentiated cells – resulted in the preparation of an enriched population to study the mechanism and kinetics of megakaryocytic differentiation. A short exposure to diosgenin was able to induce complete differentiation leading to maximal maturation which ended naturally after 192 h incubation without the influence of a secondary effect of diosgenin. The study of isolated undifferentiated cells also showed that no resistance to diosgenin was observed. This result suggested different sensitivities to differentiation induction, and SdFFF cell separation would be of great interest to explore this phenomena.