Immortalized neural stem cells from human embryonic telencephalon, HB1.F3 cells, are capable of differentiating into either neurons or glial cells in vitro and in vivo, and may be an ideal vehicle for cell replacement for neurodegenerative disease like glaucoma. We therefore examined whether intravitreally engrafted HB1.F3 cells can incorporate into the retina of glaucoma rat model eyes underwent acute ischemia-reperfusion injury. As a control, the cells were also transplanted into normal rat eyes. Engrafted HB1.F3 cells survived at least up to 2 weeks following intravitreal transplantation into both normal rat eyes and glaucoma model eyes with acute ischemic reperfusion insult. However, survived HB1.F3 cells migrate toward the inner retina of the eyes only with ischemic insult. In addition, some incorporated cells showed a morphologic differentiation into neuron with neurites. These results illustrate not only the survival of multipotent HB1.F3 cells in eyes but also the ability of these engrafted cells migrating into anatomically appropriate location to replace damaged neurons in glaucoma.