Embryonic Stem Cell Protocols

Takayuki Asano, Kyoko Sasaki, Yoshihiro Kitano, Keiji Terao, Yutaka Hanazono, Asano, Takayuki, Sasaki, Kyoko, Kitano, Yoshihiro, Terao, Keiji, Hanazono, Yutaka

To achieve human embryonic stem (ES) cell-based transplantation therapies, allogeneic transplantation models of nonhuman primates would be particularly useful. In this chapter, we describe an example of this model. We prepared cynomolgus ES cells genetically marked with the green fluorescent protein. The cells were transplanted into the allogeneic fetus because the fetus is immunologically premature and does not induce immune responses to transplanted cells. In addition, fetal tissue compartments are rapidly expanding, presumably providing space for engraftment. At 3 mo posttransplantation, a fluorescent teratoma, obviously derived from transplanted ES cells, was found in the fetus. However, transplanted cell progeny were also detected (approx 1%) in multiple fetal tissues. The cells were solitary and indistinguishable from surrounding host cells as assessed by in situ polymerase chain reaction. Transplanted cynomolgus ES cells can engraft in allogeneic fetuses. The cells will, however, form a tumor if they "leak" into an improper space, such as the thoracic cavity.