Abstract
Background aims
Amniotic fluid (AF) contains stem cells with high proliferative and differentiative potential that might be an attractive source of multipotent stem cells. We investigated whether human AF contains mesenchymal stem cells (MSC) and evaluated their phenotypic characteristics and differentiation potential in vitro.
Methods
AF was harvested during routine pre-natal amniocentesis at 14–16 weeks of pregnancy. AF sample pellets were plated in α-minimum essential medium (MEM) with 10% fetal bovine serum (FBS). We evaluated cellular growth, immunophenotype, stemness markers and differentiative potential during in vitro expansion. Neural progenitor maintenance medium (NPMM), a medium normally used for the growth and maintenance of neural stem cells, containing hFGF, hEGF and NSF-1, was used for neural induction.
Results
Twenty-seven AF samples were collected and primary cells, obtained from samples containing more than 6 mL AF, had MSC characteristics. AF MSC showed high proliferative potential, were positive for CD90, CD105, CD29, CD44, CD73 and CD166, showed Oct-4 and Nanog molecular and protein expression, and differentiated into osteoblasts, adypocytes and chondrocytes. The NPMM-cultured cells expressed neural markers and increased Na+ channel density and channel inactivation rate, making the tetrodotoxin (TTX)-sensitive channels more kinetically similar to native neuronal voltage-gated Na+ channels.
Conclusions
These data suggest that AF is an important multipotent stem cell source with a high proliferative potential able to originate potential precursors of functional neurons.
Acknowledgements
We are grateful to Dr Simona Sdei for amniotic fluid collection during routine amniocentesis performed in the Department of Obstetrics and Gynaecology, S. Anna Obstetric and Gynaecologic Hospital in Turin, Dr Andrea Marcantoni for helping with the electrophysiologic recordings, and Andrew Martin Garvey BA(Hons) LTCL for patiently revising our paper. This work was supported by ADISCO (Associazione Donatrici Italiane di Cordone Ombelicale) and Compagnia di San Paolo, Turin.
Declaration of interest: The authors report no confl icts of interest. The authors alone are responsible for the content and writing of the paper.