title: Gravitational field-flow fractionation of human hemopoietic stem cells.
abstract: New cell sorting methodologies, which are simple, fast, non-invasive, and able to isolate homogeneous cell populations, are needed for applications ranging from gene expression analysis to cell-based therapy. In particular, in the forefront of stem cell isolation, progenitor cells have to be separated under mild experimental conditions from complex heterogeneous mixtures prepared from human tissues. Most of the methodologies now employed make use of immunological markers. However, it is widely acknowledged that specific markers for pluripotent stem cells are not as yet available, and cell labelling may interfere with the differentiation process. This work presents for the first time gravitational field-flow fractionation (GrFFF), as a tool for tag-less, direct selection of human hematopoietic stem and progenitor cells from cell samples obtained by peripheral blood aphaeresis. These cells are responsible to repopulate the hemopoietic system and they are used in transplantation therapies. Blood aphaeresis sample were injected into a GrFFF system and collected fractions were characterized by flow cytometry for CD34 and CD45 expression, and then tested for viability and multi-differentiation potential. The developed GrFFF method allowed obtaining high enrichment levels of viable, multi-potent hematopoietic stem cells in specific fraction and it showed to fulfil major requirements of analytical performance, such as selectivity and reproducibility of the fractionation process and high sample recovery.
authors: Department of Chemistry G. Ciamician, Via Selmi 2, I-40126 Bologna, Italy. barbara.roda@unibo.it
authors: Barbara
authors: B
authors: Roda
authors: Pierluigi
authors: P
authors: Reschiglian
authors: Francesco
authors: F
authors: Alviano
authors: Giacomo
authors: G
authors: Lanzoni
authors: Gian Paolo
authors: GP
authors: Bagnara
authors: Francesca
authors: F
authors: Ricci
authors: Marina
authors: M
authors: Buzzi
authors: Pier Luigi
authors: PL
authors: Tazzari
authors: Pasqualepaolo
authors: P
authors: Pagliaro
authors: Elisa
authors: E
authors: Michelini
authors: Aldo
authors: A
authors: Roda
mesh_terms: D018952:Antigens, CD34 / Q000737:chemistry; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D031521:Fractionation, Field Flow / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology*; D006801:Humans; D017493:Leukocyte Common Antigens / Q000737:chemistry
chemical_list: D018952:Antigens, CD34; D017493:Leukocyte Common Antigens