Human lymphocyte sorting by gravitational field-flow fractionation.
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.
Roda
Barbara
B
Department of Chemistry "G. Ciamician", University of Bologna, Via Selmi 2, 40126, Bologna, Italy. barbara.roda@unibo.it
Reschiglian
Pierluigi
P
Zattoni
Andrea
A
Tazzari
Pier Luigi
PL
Buzzi
Marina
M
Ricci
Francesca
F
Bontadini
Andrea
A
D018941:Antigens, CD19 / Q000737:chemistry; D001402:B-Lymphocytes / Q000166:cytology* / Q000473:pathology; D002051:Burkitt Lymphoma / Q000473:pathology*; D045744:Cell Line, Tumor; D002470:Cell Survival; D005434:Flow Cytometry; D031521:Fractionation, Field Flow / Q000379:methods*; D006112:Gravitation; D006801:Humans; D017493:Leukocyte Common Antigens / Q000737:chemistry; D013601:T-Lymphocytes / Q000166:cytology* / Q000473:pathology
D018941:Antigens, CD19; D017493:Leukocyte Common Antigens