title: Routine clonal expansion of mesenchymal stem cells derived from amniotic fluid for perinatal applications.
abstract: BACKGROUND
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Development and Regeneration, KU Leuven, Leuven, Belgium.
authors.forename: Silvia
authors.initials: S
authors.lastname: Zia
authors.forename: Jaan
authors.initials: J
authors.lastname: Toelen
authors.forename: Marina
authors.initials: M
authors.lastname: Mori da Cunha
authors.forename: Philip
authors.initials: P
authors.lastname: Dekoninck
authors.forename: Paolo
authors.initials: P
authors.lastname: de Coppi
authors.forename: Jan
authors.initials: J
authors.lastname: Deprest
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology / Q000502:physiology; D005260:Female; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology / Q000637:transplantation; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*