Tunicamycin reversibly inhibits the terminal differentiation of teratocarcinoma stem cells to endoderm.

The differentiation of aggregates of certain teratocarcinoma stem cell lines begins with the formation of an outer layer of primary endoderm cells characterized by the production of plasminogen activator and the absence of histochemically detectable alkaline phosphatase activity. After several days of culture these outer cells develop into a mixture of two types of terminally differentiated endoderm: parietal endoderm which produces a thick layer of underlying basement membrane and visceral endoderm which produces alpha-fetoprotein (AFP). We report here that in the presence of tunicamycin, a drug that inhibits glycosylation of N-asparagine linked glycoproteins, a primary endoderm-like cell is formed which is alkaline phosphatase negative and plasminogen activator positive. However, terminal differentiation of these cells is inhibited as manifested by the lack of accumulation of a thick basement membrane and the absence of immunologically detected AFP. Such inhibition is reversible following removal of the tunicamycin. Terminal differentiation of endoderm depends, therefore, upon N-asparagine linked glycoproteins.