Human lymphocyte sorting by gravitational field-flow fractionation
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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.
KeywordsGravitational field-flow fractionation Cell sorting Lymphocytes
This work was supported by the Ministero Italiano dell’Istruzione, dell’Università e della Ricerca.
- 2.Salzman HM, Singham SB, Johnston RG, Bohren CF (1989) In: Melamed M, Lindmo T, Mendelshon ML (eds) Flow cytometry and sorting, 2nd edn. Wiley, New York, p 81Google Scholar
- 5.Ratanathanawongs-Williams SK (2000) In: Schimpf ME, Caldwell KC, Giddings JC (eds) Field-flow fractionation handbook, chap 17. Wiley-Interscience, New YorkGoogle Scholar
- 6.Lucas A, Lepage F, Cardot PJP (2000) In: Schimpf ME, Caldwell KC, Giddings JC (eds) Field-flow fractionation handbook, chap 29. Wiley-Interscience, New YorkGoogle Scholar
- 8.Caldwell KD, Cheng ZQ, Hradecky P, Giddings JC (1984) Cell Biophys 6:233–251Google Scholar
- 9.Battu S, Elyaman W, Hugon J, Cardot PJP (2001) Biochim Biophys Acta 1528:89–96Google Scholar
- 25.Biddison WE (2001) Curr Protoc Cell Biol Chap 2:Unit 2.2Google Scholar
- 27.Temmink JHM, Collard JG, Roosien J, Van den Bosch JF (1976) J Cell Sci 21:563–578Google Scholar
- 28.Melucci D, Roda B, Zattoni A, Casolari S, Reschiglian P, Roda A (2004) J Chromatogr A 1056:229–236Google Scholar
- 33.Epstein MA, Achong BG, Barr YM, Zajac B, Henle G, Henle W (1967) J Natl Cancer Inst 37:547–559Google Scholar