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Human lymphocyte sorting by gravitational field-flow fractionation

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Abstract

Interest in biological studies on various cell types for many biomedical applications, from research to patient treatments, is constantly increasing. The ability to discriminate (sort) and/or quantify distinct subpopulations of cells has become increasingly important. For instance, not only detection but also the highest depletion of neoplastic cells from normal cells is an important requisite in the autologous transplantation of lymphocytes for blood cancer treatments. In this work, gravitational field-flow fractionation (GrFFF) is shown to be effective for sorting a heterogeneous mixture of human, living lymphocytes constituted of neoplastic B cells from a Burkitt lymphoma cell line and healthy T and B lymphocytes from blood samples. GrFFF does not require the use of fluorescent immunotags for sorting cells, and the sorted cells can be collected for their further characterization. Flow cytometry was used to assess the viability of the cells collected, and to evaluate the cell fractionation achieved. A low amount of neoplastic B lymphocytes (less than 2%) was found in a specific fraction obtained by GrFFF. The high depletion from neoplastic cells (more than 98%) was confirmed by a clonogenicity test.

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Acknowledgement

This work was supported by the Ministero Italiano dell’Istruzione, dell’Università e della Ricerca.

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Authors and Affiliations

  1. Department of Chemistry “G. Ciamician”, University of Bologna, Via Selmi 2, 40126, Bologna, Italy

    Barbara Roda, Pierluigi Reschiglian & Andrea Zattoni

  2. National Institute of Biostructure and Biosystems, Interuniversity Consortium, Rome, Italy

    Barbara Roda, Pierluigi Reschiglian & Andrea Zattoni

  3. Service of Transfusion Medicine, S. Orsola-Malpighi Hospital, 40138, Bologna, Italy

    Pier Luigi Tazzari, Marina Buzzi, Francesca Ricci & Andrea Bontadini

Authors
  1. Barbara Roda
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  2. Pierluigi Reschiglian
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  3. Andrea Zattoni
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  4. Pier Luigi Tazzari
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  5. Marina Buzzi
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  6. Francesca Ricci
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  7. Andrea Bontadini
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Correspondence to Barbara Roda.

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Roda, B., Reschiglian, P., Zattoni, A. et al. Human lymphocyte sorting by gravitational field-flow fractionation. Anal Bioanal Chem 392, 137–145 (2008). https://doi.org/10.1007/s00216-008-2271-x

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  • DOI: https://doi.org/10.1007/s00216-008-2271-x

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