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Multipotent mesenchymal stromal cells from amniotic fluid: solid perspectives for clinical application

¹ Centro Cellule Staminali e Terapia Cellulare, Ospedale San Martino, Genova
² Laboratorio di Biologia Cardiaca e Vascolare, Clinica delle Malattie Cardiovascolari, DIMI, Università di Genova
³ Unità di Neuroimmunologia, Dipartimento di Neuroscienze Oftalmologia e Genetica and
⁴ Centro di Eccellenza per la Ricerca Biomedica, Università di Genova
⁵ Dipartimento di Scienze Genetiche, Perinatali e Ginecologiche, Ospedale Galliera, Genova
⁶ Divisione di Anatomia Patologica, Ospedale San Martino and
⁷ Dipartimento di Patologia Umana, Università di Messina, Italy

Correspondence: Francesco Frassoni, Centro Cellule Staminali e Terapia Cellulare, Ospedale San Martino, L.go R. Benzi 10, 16132 Genova Italy. E-Mail: francesco.frassoni{at}hsanmartino.it

Background: Mesenchymal stromal cells are multipotent cells considered to be of great promise for use in regenerative medicine. However, the cell dose may be a critical factor in many clinical conditions and the yield resulting from the ex vivo expansion of mesenchymal stromal cells derived from bone marrow may be insufficient. Thus, alternative sources of mesenchymal stromal cells need to be explored. In this study, mesenchymal stromal cells were successfully isolated from second trimester amniotic fluid and analyzed for chromosomal stability to validate their safety for potential utilization as a cell therapy product.

Design and Methods: Mesenchymal stromal cells were expanded up to the sixth passage starting from amniotic fluid using different culture conditions to optimize large-scale production.

Results: The highest number of mesenchymal stromal cells derived from amniotic fluid was reached at a low plating density; in these conditions the expansion of mesenchymal stromal cells from amniotic fluid was significantly greater than that of adult bone marrow-derived mesenchymal stromal cells. Mesenchymal stromal cells from amniotic fluid represent a relatively homogeneous population of immature cells with immunosuppressive properties and extensive proliferative potential. Despite their high proliferative capacity in culture, we did not observe any karyotypic abnormalities or transformation potential in vitro nor any tumorigenic effect in vivo.

Conclusions: Fetal mesenchymal stromal cells can be extensively expanded from amniotic fluid, showing no karyotypic abnormalities or transformation potential in vitro and no tumorigenic effect in vivo. They represent a relatively homogeneous population of immature mesenchymal stromal cells with long telomeres, immunosuppressive properties and extensive proliferative potential. Our results indicate that amniotic fluid represents a rich source of mesenchymal stromal cells suitable for banking to be used when large amounts of cells are required.

Key words: amniotic fluid, fetal stem cells, adult mesenchymal stromal cells, karyotype.