Introduction: Amniotic fluid (AF) contains stem cells which, due to their ontogenetic origin, have a high proliferative and differentiative potential. The aim of our work was to set up a method for mesenchymal stem cells (MSCs) isolation and expansion and to evaluate their immunophenotype characteristics and differentiation capacity. Methods: AF was harvested from women undergoing aminocentesis for routine prenatal diagnosis at 14–16 weeks of pregnancy. AF samples were centrifuged and the resulting pellets were plated?-Mem with 10% Fetal Bovine Serum. Cellular growth, immunophenotype, stemness markers, telomere length and differentiative potential were evaluated during the in vitro expansion. Neural Progenitor Maintenance Medium (NPMM, Lonza) was used for neural induction. Results: Twenty six primary cellular cultures were obtained from 28 samples. The cells obtained from the samples containing more than 6 ml of AF, had fibroblastic aspects with a high proliferative potential and were positive for CD90, CD105, CD29, CD44, CD166. The expansion did not induce a significant telomere length shortening and the molecular analysis showed the expression of Oct-4, Nanog and Rex-1. Moreover, the MSCs isolated from AF differentiated in osteoblasts, adypocytes and chondrocytes. The cells cultured in NPMM expressed neural markers and functionally active voltage-dependent Na1 and Ca21 channels. Conclusion: All these data suggested that AF is an important multipotent stem cell source with a high proliferative potential able to transdifferentiate in cells with neuronal characteristics.
Mesenchymal stem cells isolated from amniotic fluid differentiate in neural cells
MARESCHI, Katia;COMUNANZA, Valentina;FERRERO, Ivana;CARBONE, Emilio;Fagioli F.
2008-01-01
Abstract
Introduction: Amniotic fluid (AF) contains stem cells which, due to their ontogenetic origin, have a high proliferative and differentiative potential. The aim of our work was to set up a method for mesenchymal stem cells (MSCs) isolation and expansion and to evaluate their immunophenotype characteristics and differentiation capacity. Methods: AF was harvested from women undergoing aminocentesis for routine prenatal diagnosis at 14–16 weeks of pregnancy. AF samples were centrifuged and the resulting pellets were plated?-Mem with 10% Fetal Bovine Serum. Cellular growth, immunophenotype, stemness markers, telomere length and differentiative potential were evaluated during the in vitro expansion. Neural Progenitor Maintenance Medium (NPMM, Lonza) was used for neural induction. Results: Twenty six primary cellular cultures were obtained from 28 samples. The cells obtained from the samples containing more than 6 ml of AF, had fibroblastic aspects with a high proliferative potential and were positive for CD90, CD105, CD29, CD44, CD166. The expansion did not induce a significant telomere length shortening and the molecular analysis showed the expression of Oct-4, Nanog and Rex-1. Moreover, the MSCs isolated from AF differentiated in osteoblasts, adypocytes and chondrocytes. The cells cultured in NPMM expressed neural markers and functionally active voltage-dependent Na1 and Ca21 channels. Conclusion: All these data suggested that AF is an important multipotent stem cell source with a high proliferative potential able to transdifferentiate in cells with neuronal characteristics.
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