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A simple approach for mouse embryonic stem cells isolation and differentiation inducing embryoid body formation
Author(s) -
Fagundez Carol B.,
Loresi Mónica A.,
Quintana Ojea Marcos E.,
Delcourt Stella M.,
Testa Roberto,
Gogorza Sebastián J.,
Argibay Pablo F.
Publication year - 2009
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1016/j.cellbi.2009.08.001
Subject(s) - embryoid body , embryonic stem cell , kosr , stem cell , blastocyst , biology , microbiology and biotechnology , alkaline phosphatase , fetal bovine serum , fetus , immunology , andrology , cell culture , embryo , adult stem cell , embryogenesis , biochemistry , genetics , medicine , enzyme , pregnancy , gene
Stem cells were derived from hatched blastocyst‐stage mouse embryos of the C57BL/6 strain employing a knockout serum replacement instead of the traditional fetal calf serum, thereby avoiding the use of immunosurgery. Although fetal calf serum was not good for isolation of stem cells, a combination of this serum plus knockout serum increased the expansion rate of the cell culture. The derived cells were capable of maintaining an undifferentiated state during several passages, as demonstrated by the presence of alkaline phosphatase activity, stage‐specific embryonic antigen 1 (SSEA‐1), and octamer binding protein 4 (Oct‐4). Suspension culture in bacteriological dishes gave better results than the hanging drop method for differentiation by means of embryoid body formation. Mouse embryonic stem cells showed spontaneous differentiation into derivatives of the 3 germ layers in culture media supplemented with fetal calf serum but not with knockout serum.