The use of stem cells in regenerative medicine and cell-based therapies offers immense potential in diseases which have currently no treatment such as Duchenne muscular dystrophy. A possible limitation to the use of CD133+ for a therapeutic application is the relatively low number of cells that can be recovered from peripheral blood mononuclear cells. The goal of ex vivo expansion is to induce proliferation of CD133+ cells while maintaining their primary functional characteristic, namely, their ability to engraft and differentiate. In this work, we expanded in culture stem cells with different combinations of cytokines, to explore possibility of ex vivo expansion of CD133+ cells from peripheral blood. All experiments cultured with different combinations of cytokines supported the growth of CD133+ cells compared with the group without cytokines, but only a cocktail including SCF, bFGF, EGF, VEGF, LIF, TEPA, IL6 showed a significant expansion of stem cells in culture. In this condition we were able to expand the CD133+ cells for more than 50 passages over a 2-month period and we observed no indication of replicative senescence or significant changes in cellular division time during expansion period. Proliferating cells still had the capacity to form hematopoietic and endothelial colonies in semisolid media. Furthermore, we showed that expanded populations of CD133+ cells derived from blood maintain the capacity to differentiate into myogenic cells in vitro and in vivo. Human circulating CD133+ cells were also cultured at 5- or 20-percent oxygen in liquid culture in presence of the better cocktail of cytokines and we analysed and compared their expansion capacity and their vitality. The total number of cells increased 6-fold at 5-percent oxygen and could result in a better maintenance of the balance between primitive progenitor cell renewal and clonogenic progenitor expansion, thus representing a tool of remarkable therapeutic interest.

Ex vivo expansion of human circulanting CD133+promising tool for cell-based therapeutic approaches in muscular dystrophy / M. Belicchi, M. Meregalli, A. Farini, A. Cattaneo, C. Marchesei, L. Porretti, N. Bresolin, Y. Torrente. - In: NEUROMUSCULAR DISORDERS. - ISSN 0960-8966. - 17:9-10(2007 Oct), pp. 875-876. ((Intervento presentato al 12. convegno International congress of the world muscle society tenutosi a Taormina nel 2007 [10.1016/j.nmd.2007.06.381].

Ex vivo expansion of human circulanting CD133+promising tool for cell-based therapeutic approaches in muscular dystrophy

M. Belicchi
Primo
;
M. Meregalli;A. Farini;A. Cattaneo;N. Bresolin
Penultimo
;
Y. Torrente
Ultimo
2007-10

Abstract

The use of stem cells in regenerative medicine and cell-based therapies offers immense potential in diseases which have currently no treatment such as Duchenne muscular dystrophy. A possible limitation to the use of CD133+ for a therapeutic application is the relatively low number of cells that can be recovered from peripheral blood mononuclear cells. The goal of ex vivo expansion is to induce proliferation of CD133+ cells while maintaining their primary functional characteristic, namely, their ability to engraft and differentiate. In this work, we expanded in culture stem cells with different combinations of cytokines, to explore possibility of ex vivo expansion of CD133+ cells from peripheral blood. All experiments cultured with different combinations of cytokines supported the growth of CD133+ cells compared with the group without cytokines, but only a cocktail including SCF, bFGF, EGF, VEGF, LIF, TEPA, IL6 showed a significant expansion of stem cells in culture. In this condition we were able to expand the CD133+ cells for more than 50 passages over a 2-month period and we observed no indication of replicative senescence or significant changes in cellular division time during expansion period. Proliferating cells still had the capacity to form hematopoietic and endothelial colonies in semisolid media. Furthermore, we showed that expanded populations of CD133+ cells derived from blood maintain the capacity to differentiate into myogenic cells in vitro and in vivo. Human circulating CD133+ cells were also cultured at 5- or 20-percent oxygen in liquid culture in presence of the better cocktail of cytokines and we analysed and compared their expansion capacity and their vitality. The total number of cells increased 6-fold at 5-percent oxygen and could result in a better maintenance of the balance between primitive progenitor cell renewal and clonogenic progenitor expansion, thus representing a tool of remarkable therapeutic interest.
Settore MED/26 - Neurologia
World Muscle society
Article (author)
File in questo prodotto:
File Dimensione Formato  
1-s2.0-S0960896607005688-main.pdf

accesso riservato

Tipologia: Publisher's version/PDF
Dimensione 64.2 kB
Formato Adobe PDF
64.2 kB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/44657
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? 0
social impact