TY - JOUR
T1 - Characterization of pluripotent stem cells
AU - Martí, Mercè
AU - Mulero, Lola
AU - Pardo, Cristina
AU - Morera, Cristina
AU - Carrió, Meritxell
AU - Laricchia-Robbio, Leopoldo
AU - Esteban, Concepcion Rodriguez
AU - Belmonte, Juan Carlos Izpisua
PY - 2013/2
Y1 - 2013/2
N2 - Characterization of pluripotent stem cells is required for the registration of stem cell lines and allows for an impartial and objective comparison of the results obtained when generating multiple lines. It is therefore crucial to establish specific, fast and reliable protocols to detect the hallmarks of pluripotency. Such protocols should include immunocytochemistry (takes 2 d), identification of the three germ layers in in vitro-derived embryoid bodies by immunocytochemistry (immunodetection takes 3 d) and detection of differentiation markers in in vivo-generated teratomas by immunohistochemistry (differentiation marker detection takes 4 d). Standardization of the immunodetection protocols used ensures minimum variations owing to the source, the animal species, the endogenous fluorescence or the inability to collect large amounts of cells, thereby yielding results as fast as possible without loss of quality. This protocol provides a description of all the immunodetection procedures necessary to characterize mouse and human stem cell lines in different circumstances.
AB - Characterization of pluripotent stem cells is required for the registration of stem cell lines and allows for an impartial and objective comparison of the results obtained when generating multiple lines. It is therefore crucial to establish specific, fast and reliable protocols to detect the hallmarks of pluripotency. Such protocols should include immunocytochemistry (takes 2 d), identification of the three germ layers in in vitro-derived embryoid bodies by immunocytochemistry (immunodetection takes 3 d) and detection of differentiation markers in in vivo-generated teratomas by immunohistochemistry (differentiation marker detection takes 4 d). Standardization of the immunodetection protocols used ensures minimum variations owing to the source, the animal species, the endogenous fluorescence or the inability to collect large amounts of cells, thereby yielding results as fast as possible without loss of quality. This protocol provides a description of all the immunodetection procedures necessary to characterize mouse and human stem cell lines in different circumstances.
UR - http://www.scopus.com/inward/record.url?scp=84873373021&partnerID=8YFLogxK
U2 - 10.1038/nprot.2012.154
DO - 10.1038/nprot.2012.154
M3 - Article
C2 - 23306458
AN - SCOPUS:84873373021
SN - 1754-2189
VL - 8
SP - 223
EP - 253
JO - Nature Protocols
JF - Nature Protocols
IS - 2
ER -