Routine clonal expansion of mesenchymal stem cells derived from amniotic fluid for perinatal applications

• Published in: Prenatal diagnosis Vol. 33; no. 10; pp. 921 - 928
• Main Authors: Zia, Silvia, Toelen, Jaan, Mori da Cunha, Marina, DeKoninck, Philip, de Coppi, Paolo, Deprest, Jan
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.10.2013; Wiley Subscription Services, Inc
• Subjects: Amniotic Fluid - cytology; Animals; Animals, Newborn; Cell Culture Techniques - methods; Cell Proliferation; Cells, Cultured; Clone Cells - cytology; Clone Cells - physiology; Female; Humans; Mesenchymal Stem Cell Transplantation; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - physiology; Mice; Mice, SCID; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - physiology; Pluripotent Stem Cells - transplantation; Pregnancy; Prenatal Diagnosis - methods
• ISSN: 0197-3851; 1097-0223
• DOI: 10.1002/pd.4162

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. © 2013 John Wiley & Sons, Ltd. What's already known about this topic? Amniotic fluid‐derived mesenchymal stem cells have the unique advantage of extensive self‐renewal in vitro allowing for long‐term expansion, full characterization, and cryopreservation. What does this study add? Isolation procedures were possible in samples derived from mid‐gestation amniocentesis samples. The isolation procedure was reproducible and applicable in preclinical research. Amniotic fluid‐derived mesenchymal stem cells can be genetically modified with a lentiviral vector and be used for in vivo cell therapy experiments.