This study was supported in part by the Cord Blood Stem Cell Project (Fondazione Cassa di Risparmio di Roma, Rome, Italy) and by Ministero dell’Università e della Ricerca Scientifica e Tecnologica (MURST, Rome, Italy).
Clinical isolation and functional characterization of cord blood CD133+ hematopoietic progenitor cells†
Article first published online: 28 JUN 2004
Volume 44, Issue 7, pages 1087–1097, July 2004
How to Cite
Bonanno, G., Perillo, A., Rutella, S., De Ritis, D. G., Mariotti, A., Marone, M., Meoni, F., Scambia, G., Leone, G., Mancuso, S. and Pierelli, L. (2004), Clinical isolation and functional characterization of cord blood CD133+ hematopoietic progenitor cells. Transfusion, 44: 1087–1097. doi: 10.1111/j.1537-2995.2004.03252.x
- Issue published online: 28 JUN 2004
- Article first published online: 28 JUN 2004
- Received for publication July 28, 2003; revision received February 20, 2004, and accepted February 23, 2004.
BACKGROUND: Human cord blood is a relevant source of CD133+ HPCs. Clinical-scale isolation of human umbi-lical cord blood (UCB) CD133+ HPCs using immunomag-netic microbeads and the CliniMACS clinical cell isolator is reported. CD133+ HPCs isolated after large-scale pro-cessing were functionally characterized.
STUDY DESIGN AND METHODS: Closed disposable sets were used to process nine different samples of RBC-reduced UCB nucleated cells. In-vitro hematopoietic assays and human xenografts in NOD/SCID mice were performed to assess the functional properties of isolated CD133+ cells. Different mixtures of human cytokines were tested for the ability to expand nascent CD133+ HPCs. Furthermore, freshly isolated CD133+ cells were conditioned in culture medium specifically tested to support in-vitro myogenesis or osteogenesis.
RESULTS: Isolation procedures yielded the recovery of an average of 2.53 × 106 CD133+ HPCs with a mean recovery of 96 percent (referred to as RBC-reduced samples) and a final sample purity of 82 percent. Purified CD133+ cells had high cloning efficiency, had relevant long-term activity, and were capable of repopulating irradiated NOD/SCID mice. In 10-day stroma-free cultures, a 2-fold and 8.3-fold expansion of colony-forming cells (CFCs) and extended long-term culture-initiating cells, respectively, was obtained. Freshly isolated CD133+ cells differen-tiated into large nucleated cells expressing either myosin D or osteopontin (as revealed by RT-PCR and immuno-cytochemistry), with a protein/mRNA expression compar-able to or even higher than that observed in UCB CD133– nucleated cells in identical culture conditions.
CONCLUSION: Collectively, clinical-scale isolation of UCB CD133+ cells provides a relevant amount of primi-tive HPCs with high hematopoietic activity and in-vitro mesenchymal potential.