Embryoid body in vitro formation from human amniotic fluid stem cells (AFSCs): an ultrastructural study (Abstract in rivista)

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  • Embryoid body in vitro formation from human amniotic fluid stem cells (AFSCs): an ultrastructural study (Abstract in rivista) (literal)
Anno
  • 2013-01-01T00:00:00+01:00 (literal)
Alternative label
  • Maria Antonietta Centurione1,2, Ivana Antonucci2,3, Lucia Centurione2,4, Liborio Stuppia2,3 and Roberta Di Pietro2,4 (2013)
    Embryoid body in vitro formation from human amniotic fluid stem cells (AFSCs): an ultrastructural study
    (literal)
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  • Maria Antonietta Centurione1,2, Ivana Antonucci2,3, Lucia Centurione2,4, Liborio Stuppia2,3 and Roberta Di Pietro2,4 (literal)
Pagina inizio
  • 53 (literal)
Pagina fine
  • 53 (literal)
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  • N. 2 (Supplement) (literal)
Rivista
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  • 1 (literal)
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  • 118 (literal)
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  • 1 Institute of Molecular Genetics, National Research Council-Pavia, Section of Chieti, Italy 2 Ageing Research Center, CeSI, G. d'Annunzio University Foundation, StemTeCh Group, Chieti, Italy 3 Laboratory of Molecular Genetics, Department of Psychological, Humanistic and Territorial Sciences, G. d'Annunzio University of Chieti-Pescara, Italy 4 Department of Medicine and Ageing Sciences, G. d'Annunzio University of Chieti-Pescara, Italy (literal)
Titolo
  • Embryoid body in vitro formation from human amniotic fluid stem cells (AFSCs): an ultrastructural study (literal)
Abstract
  • Amniotic fluid stem cells (AFSCs) harbour the potential to differentiate into cells of any of the three germ layers and to form embryoid bodies (EBs) without induc- ing teratoma formation (De Coppi et al., 2007; Antonucci et al., 2012). However, no previous findings have been reported regarding embryoid body in vitro development and ultrastructural organization. Thus, this was the aim of our study. Amniotic fluid samples were obtained from women undergoing amniocentesis for prenatal diagno- sis at 16-19 weeks of pregnancy after written informed consent and the local ethical committee approval. Human AFSCs were cultured up to the 8th passage and analysed with RT-PCR for the expression of pluripotency markers. Some cellular pools were cultured in suspension in uncoated Petri dishes (hanging drop method) to obtain EB formation. After 5 days of culture, the appearance of EBs of different size was observed with phase contrast microscopy and monitored up to 10-15 days of culture. In parallel, EB standard embedding in paraffin for light microscopy or in epoxy resin for transmission electron microscopy was performed. RT-PCR analysis revealed the presence of classical markers of pluripotency (OCT4, NANOG, SOX2) in AFSCs at the 2th-8th passage, whereas specific markers of the three embryonic germ layers were detected in EB specimens. Morphological assays of three-dimensional aggregates demonstrated the presence of solid structures only at the beginning of the culture whereas signs of apoptotic cell death accompanied by the secretion of an amorphous substance were soon detected. These features were preliminary to the development, at later culture time intervals, of an inner hollow cavity surrounded by a crown of flat cells displaying a number of electron dense granules and highly resembling tro- phoblastic cells. (literal)
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