Open AccessTeratoma Formation: A Tool for Monitoring Pluripotency in Stem Cell Research
Author(s) -
Nelakanti Raman V.,
Kooreman Nigel G.,
Wu Joseph C.
Publication year - 2015
Publication title -
current protocols in stem cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.658
H-Index - 28
eISSN - 1938-8969
pISSN - 1941-7322
DOI - 10.1002/9780470151808.sc04a08s32
Subject(s) - ectoderm , endoderm , biology , germ layer , teratoma , mesoderm , embryonic stem cell , induced pluripotent stem cell , microbiology and biotechnology , stem cell , pathology , anatomy , embryogenesis , embryo , medicine , genetics , gene
This unit describes protocols for evaluating the pluripotency of embryonic and induced pluripotent stem cells using a teratoma formation assay. Cells are prepared for injection and transplanted into immunodeficient mice at the gastrocnemius muscle, a site well suited for teratoma growth and surgical access. Teratomas that form from the cell transplants are explanted, fixed in paraformaldehyde, and embedded in paraffin. These preserved samples are sectioned, stained, and analyzed. Pluripotency of a cell line is confirmed by whether the teratoma contains tissues derived from each of the embryonic germ layers: endoderm, mesoderm, and ectoderm. Alternatively, explanted and fixed teratomas can be cryopreserved for immunohistochemistry, which allows for more detailed identification of specific tissue types present in the samples. © 2015 by John Wiley & Sons, Inc.