Implantation of human MSCs promoted proliferation, migration, and survival of mouse NSCs. (A) Real-time RT-PCR assays for PCNA. Error bars indicate standard errors (n = 4). A significant increase (*, p < 0.05) over vehicle-injected controls was seen at 1 and 3 days after implantation. (B) Real-time RT-PCR assays for PCNA after control implantation of nonviable MSCs. There was no increase in the number of proliferating cells at 3 days after implantation (n = 4). (C) Increased proliferation was observed in the implanted hippocampus compared with the vehicle-injected control at 7 days after implantation. MSCs (anti-HNA, red) remained localized to the DG, whereas proliferating cells (anti-PCNA, green) were seen in the stratum radiatum, CA1 pyramidal layer, and stratum oriens of the hippocampus. (Scale bar, 100 μm.) (D) Experimental schedule for labeling of newly generated cells. Dashed arrows indicate MSC implantation, solid arrows indicate BrdUrd injections, and arrowheads indicate animal sacrifice. (E) Immunohistochemistry of the hippocampus at 7 or 30 days after implantation. Cells were no longer proliferating by 30 days after implantation, but the newly generated cells (anti-BrdUrd, red) persisted. Proliferating cells expressed the NSC marker Sox-2 (red), and a subpopulation of newly generated cells expressed the neuronal migration marker doublecortin (green). Arrows, colabeled cells; arrowheads, singly labeled cells. (Scale bars, 25 μm.)