Gravitational field-flow fractionation of human hemopoietic stem cells

Barbara Roda, Pierluigi Reschiglian, Francesco Alviano, Giacomo Lanzoni, Gian Paolo Bagnara, Francesca Ricci, Marina Buzzi, Pier Luigi Tazzari, Pasqualepaolo Pagliaro, Elisa Michelini, Aldo Roda

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)9081-9087
Number of pages7
JournalJournal of Chromatography A
Volume1216
Issue number52
DOIs
StatePublished - Dec 25 2009
Externally publishedYes

Keywords

  • Cell sorting
  • Gravitational field-flow fractionation
  • Hematopoietic stem cells

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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