Multipotent mesenchymal stem cells from amniotic fluid originate neural precursors with functional voltage-gated sodium channels.
OBJECTIVE
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 alpha-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.
Mareschi
Katia
K
Stem Cell Transplantation and Cellular Therapy Unit, Pediatric Onco-Hematology Department, Regina Margherita Children's Hospital, Turin, Italy. Katia.mareschi@unito.it
Rustichelli
Deborah
D
Comunanza
Valentina
V
De Fazio
Roberta
R
Cravero
Cristina
C
Morterra
Giulia
G
Martinoglio
Barbara
B
Medico
Enzo
E
Carbone
Emilio
E
Benedetto
Chiara
C
Fagioli
Franca
F
D000653:Amniotic Fluid / Q000166:cytology*; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation; D048430:Cell Shape; D002478:Cells, Cultured; D003470:Culture Media; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D016130:Immunophenotyping; D015640:Ion Channel Gating; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D009474:Neurons / Q000166:cytology* / Q000378:metabolism*; D011247:Pregnancy; D015222:Sodium Channels / Q000378:metabolism*
D015415:Biomarkers; D003470:Culture Media; D015222:Sodium Channels