Abstract
INTRODUCTION: Stem cells (SCs) isolated from amniotic fluid (AF) are a promising source for autologous perinatal cell therapy. The aim of this study was to develop a routine isolation, selection, and expansion protocol of clonal SC lines from redundant clinical amniocentesis samples.
MATERIALS AND METHODS: Amniotic fluids were collected between 15 and 22 weeks of gestation, and SCs were isolated by CD117-based and mechanical selection protocols. SCs were characterized by mesenchymal SC marker expression and differentiation protocols. Cells were manipulated with a lentiviral vector system expressing the β-galactosidase reporter gene and were injected into immunodeficient newborn mouse pups. Qualitative assessment was performed to detect the infused cells after 1 week.
RESULTS: A total of 78 clonal AF SC populations were successfully isolated by mechanical selection from 21 consecutive amniocentesis samples. They were positive for mesenchymal SC cluster of differentiation markers and could be differentiated into the different lineages. SCs were stably labeled using β-galactosidase and were detected in the lungs and hearts of the neonatal mice.
CONCLUSION: We demonstrate that mesenchymal SCs can be routinely isolated and clonally expanded from mid-gestation human AF using mechanical isolation. They can easily be transduced and be tested for perinatal treatment in animal models.
| Original language | English |
|---|---|
| Pages (from-to) | 921-928 |
| Number of pages | 8 |
| Journal | Prenatal Diagnosis |
| Volume | 33 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - Oct 2013 |
| Externally published | Yes |
Bibliographical note
© 2013 John Wiley & Sons, Ltd.Funding sources:
This work was supported by grants of the European Commission in its 6th Framework Program (MEST CT2005 019707; EuroSTEC, LSHC-CT2006-037409), and the Instituut voor Wetenschap en Technologie (I.W.T.). Jaan Toelen has been supported by the Klinisch Onderzoeksfonds of the University
Hospitals Leuven. Jan Deprest is a Clinical Researcher for the Fonds Wetenschappelijk Onderzoek-Vlaanderen (1.8.012.07.N.02).
Conflicts of interest: None declared