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Testis Cancer as a model for human pluripotent stem cell differentiation.
Author(s) -
Schopperle William M,
DeWolf William C
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a873
Subject(s) - podocalyxin , induced pluripotent stem cell , cellular differentiation , stem cell , biology , microbiology and biotechnology , embryonic stem cell , biochemistry , genetics , gene , podocyte , proteinuria , kidney
Currently, there are only three sources of human pluripotent stem cells, two of which are ethically controversial sources because they are derived from human fetuses and embryos. However, the third source, embryonal carcinoma (EC), is a pluripotent stem cell derived from human germ cell tumors. All human pluripotent cells share common stem cell‐defining markers including the TRA‐1‐60 and TRA‐1‐81 antigens, which disappear from the stem cell surface upon differentiation. We have previously shown that the cell adhesion protein, podocalyxin, is the molecular carrier of the TRA‐1‐60/TRA‐1‐81 antigens on EC. In this study, we have used an EC differentiation model to study the fate of podocalyxin and its TRA‐1‐60/TRA‐1‐81 epitopes during EC differentiation. Western blots were performed on cell protein lysates and purified podocalyxin from undifferentiated EC and retinoic acid‐induced differentiated EC cells using TRA‐1‐60, TRA‐1‐81 and podocalyxin specific monoclonal antibodies. Podocalyxin is expressed as a 200 kDa protein in EC and is specific for the TRA‐1‐60 and TRA‐1‐81 antibodies. Podocalyxin expression in differentiated EC is modified; Podocalyxin is detected as a smaller protein and no longer reacts with the TRA‐1‐60/TRA‐1‐81 antibodies. These results provide the first step in determining the biochemical mechanisms of the loss of the TRA‐1‐60/TRA‐1‐81 markers during stem cell differentiation.