Pluripotent differentiation in vitro of murine ES-D3 embryonic stem cells.
Although the ES-D3 murine embryonic stem cell line was one of the first derived, little information exists on the in vitro differentiation potential of these cells. We have used immunocytochemical and flow cytometric methods to monitor ES-D3 embryoid body differentiation in vitro during a 21-d period. Spontaneous differentiation of embryoid body cells was induced by leukemia inhibitory factor withdrawal in the absence of feeder cells. The pluripotent stem cell markers Oct-3/4, SSEA-1, and EMA-1 were found to persist for at least 7 d, whereas the primitive endoderm marker cytokeratin endo-A was expressed at increasing levels from day 6. The localization of these antigens within the embryoid bodies suggested that embryonic ectoderm- and primitive endoderm-derived tissues were segregated. Localized expression of class III beta-tubulin and sarcomeric myosin also was detected, indicating that representatives of all three embryonic germ layers were present after induction of differentiation in vitro.
Toumadje
Arazdordi
A
National Stem Cell Resource, American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 20110, USA. toumadje@atcc.org
Kusumoto
Ken-Ichi
K
Parton
Angela
A
Mericko
Patricia
P
Dowell
Lori
L
Ma
Guozhong
G
Chen
Luping
L
Barnes
David W
DW
Sato
J Denry
JD
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D004622:Embryo, Mammalian; D005455:Fluorescent Antibody Technique; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D051379:Mice
D000911:Antibodies, Monoclonal; D000469:Alkaline Phosphatase