Isolation of Human Embryonic Stem Cell–Derived Teratomas for the Assessment of Pluripotency

This unit describes protocols on how to assess the developmental potency of human embryonic stem cells (hESCs) by performing xenografting into immunodeficient mice to induce teratoma formation. hESCs can be injected under the testis capsule, or alternatively into the kidney or subcutaneously. Teratomas that develop from grafted hESCs are surgically removed, fixed in formaldehyde, and paraffin embedded. The tissues in the teratoma are analyzed histologically to determine whether the hESCs are pluripotent and form tissues derived from of all three embryonic germ layers (ectoderm, mesoderm, and endoderm). Teratomas can also be fixed in Bouin's or cryosectioned for analysis, and they can be analyzed by immunohistochemistry for tissue markers. Methods for these procedures are included in this unit. Curr. Protoc. Stem Cell Biol. 3:1B.4.1-1B.4.29. © 2007 by John Wiley & Sons, Inc. Keywords: human embryonic stem cells; pluripotency; teratoma; immunodeficient mice