title: Lifespan of human amniotic fluid-derived multipotent mesenchymal stromal cells.
abstract: OBJECTIVE
  Human multipotent mesenchymal stromal cells (hMSC) have become one of the main interests in regenerative medicine because of their ability to differentiate into different lineages. Human amniotic fluid is reported to contain MSC (hAMSC) and therefore may be a useful source of cells for clinical applications. However, our understanding of the behavior of these cells in indefinite in vitro culture conditions is very limited.
  
  
  METHODS
  We systematically evaluated and characterized, throughout their whole lifespan, the expansion potential, chromosomal stability, surface and intracellular phenotype and differentiation potential of fibroblastoid hAMSC (F-type hAMSC).
  
  
  RESULTS
  Nine F-type hAMSC cultures could be expanded in in vitro culture conditions for 223.25 ± 24.44 days (mean ± SD), during which time 28.96 ± 1.5 passages were made giving rise to 54.95 ± 3.17 population doublings (PD) and an estimated number of accumulated cells of between 1.0 × 10(22) and 9.7 × 10(23), with no visible alterations in the chromosome during their lifespan. All the cultures showed unchanged percentages of strongly positive expressions of the surface markers CD29, CD44, CD73, CD90, CD95, CD105 and HLA-ABC, as well as the embryonic intracellular markers Nanog and Sox2, during their lifespan, whereas the expression of the embryonic surface markers SSEA3, SSEA4, TRA-1-60 and TRA-1-81 fell until it disappeared with progression of the culture. These cells retained their differentiation capacities to adipogenic, chondrogenic and osteogenic lineages throughout their lifespan.
  
  
  CONCLUSIONS
  F-type hAMSC exhibit reproducible biologic characteristics, confirming that these cells are ideal candidates for use in regenerative medicine.
authors.affiliation: Immunology Service, Regional University Hospital Carlos Haya, Malaga, Spain.
authors.forename: Jose Maria
authors.initials: JM
authors.lastname: Miranda-Sayago
authors.forename: Nieves
authors.initials: N
authors.lastname: Fernández-Arcas
authors.forename: Carmen
authors.initials: C
authors.lastname: Benito
authors.forename: Armando
authors.initials: A
authors.lastname: Reyes-Engel
authors.forename: Jorge
authors.initials: J
authors.lastname: Carrera
authors.forename: Antonio
authors.initials: A
authors.lastname: Alonso
mesh_terms: D017667:Adipocytes / Q000166:cytology / Q000502:physiology; D000328:Adult; D000653:Amniotic Fluid / Q000166:cytology*; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D016922:Cellular Senescence; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D039902:Multipotent Stem Cells / Q000166:cytology / Q000502:physiology*; D055746:Stage-Specific Embryonic Antigens / Q000032:analysis; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology; D055815:Young Adult
chemical_list: D055746:Stage-Specific Embryonic Antigens