13° WORLD CONGRESS ON HUMAN REPRODUCTION

Venice, March 5-7, 2009

Amniotic fluid13° WORLD CONGRESS ON HUMAN REPRODUCTION

Venice, March 5-7, 2009


Amniotic fluid mesenchymal stem cells isolation, characterization and cryopreservation.

Daniela Pecoraro1, Elena Sanna1,2, Massimiliano Manganini2, Francesca R. Grati1, Federico Maggi1,2, Giuseppe Simoni1,2

1Research and Development, TOMA Advanced Biomedical Assays S.p.A., Busto Arsizio, Varese, Italy
2Biocell Center, Busto Arsizio, Varese, Italy

ABSTRACT
Amniotic fluid draws great interest as a source of mesenchymal stem cells (AF-MSCs) for regenerative medicine. Herein we present a study aimed to characterize AF-MSCs. We analysed the chromosome stability after in vitro culture on 7 native AF samples. Afterwards on 10 morphologically homogeneous clones we evaluated i) the correlation between cell morphology and multipotent potential by investigating their time of doubling, expression of MSC markers and differentiation potential; ii) the microsatellite instability. Finally, we tested the yield of cell growth after cryopreservation on 40 native AF samples. Karyotype and microsatellite instabilities were excluded until the 5th and 18th culture passages, respectively. The 10 homogeneous clones displayed the following morphologies: 2 were fibroblastoid (F), 6 of amniotic fluid-type (AF) and 2 were epitelyoid (E). The two F clones and 2/6 AF ones demonstrated the immunophenotype, the cell growth profile and the differential potential typical of the MSCs. Finally, the cryopreservation did not adversely affect the viability and cell growth of AF cells. In conclusion, our finding indicate that: i) in addition to F-type also the AF-type cells may display a MSCs profile; ii) the integrity of the genome is preserved during in vitro cell culture: this is an important parameter for a therapeutic use of AF-MSCs; iii) cryopreservation of native AF cells do not jeopardize the cell viability. The storage of native samples without laboratory procedure/manupulation (i.e.: the culture on growth media with unknown composition) is the advisable solution to provide AF-MSCs source for future applicative therapeutic protocols on humans.