Volume 3, Issue 3, Pages 646-650 (March 2013) Induced Pluripotent Stem Cell-Derived Neural Cells Survive and Mature in the Nonhuman Primate Brain  Marina E. Emborg, Yan Liu, Jiajie Xi, Xiaoqing Zhang, Yingnan Yin, Jianfeng Lu, Valerie Joers, Christine Swanson, James E. Holden, Su-Chun Zhang  Cell Reports  Volume 3, Issue 3, Pages 646-650 (March 2013) DOI: 10.1016/j.celrep.2013.02.016 Copyright © 2013 The Authors Terms and Conditions

Cell Reports 2013 3, 646-650DOI: (10.1016/j.celrep.2013.02.016) Copyright © 2013 The Authors Terms and Conditions

Figure 1 Differentiation of Grafted Neural Progenitors (A) Schematic diagram of iPSC differentiation to neural progenitors and dopaminergic neurons. (B) Quantification of cell populations at day 42 (the day for transplantation). (C) On day 14, neuroepithelia organize into rosettes and express Pax6. By day 42, most cells express Tuj1, and some are positive for TH and GABA. (D) Six months posttransplantation, Hoechst-labeled nuclei and corresponding GFP-labeled graft in the putamen show nondisrupted cytoarchitecture of the grafted area as well as the graft and cellular projections. (E) Quantification of neurons (MAP2+), astrocytes (GFAP+), and oligodendrocytes (MBP+) among GFP+ grafted cells. (F) Colabeling of GFP with neuronal (Tuj1, MAP2, NeuN, TH, GABA) and glial (GFAP, MBP) markers in the graft. HO, Hoechst. The bar represents 50 μm. Data are presented as mean ± SE. See also Figure S1. Cell Reports 2013 3, 646-650DOI: (10.1016/j.celrep.2013.02.016) Copyright © 2013 The Authors Terms and Conditions

Figure 2 Reaction of Glia and Infiltration of Leukocytes in the Grafts Shown are representatives of grafts in the putamen revealed by GFP. These sections are colabeled with antibodies against HLA-DR (for microglia and macrophages), CD3 and CD8 (for lymphocytes), as well as GFAP (for astrocytes). Lower row is the spleen tissue sections as positive controls for HLA-DR, CD3, and CD8. HO, Hoechst. The bar represents 50 μm. See also Figure S2. Cell Reports 2013 3, 646-650DOI: (10.1016/j.celrep.2013.02.016) Copyright © 2013 The Authors Terms and Conditions

Figure S1 Generation, Genetic Labeling, and Teratoma Formation of Rhesus iPSCs, Related to Figure 1 (A) Rhesus fibroblasts. (B) An iPSC colony derived from rhesus fibroblasts. Inset shows the typical stem cell morphology. (C and D) rhesus iPSCs are positive for Sox2 (C) and Nanog (D). (E) A rhesus iPSC colony exhibit green GFP after infection with lentiviral PGK-GFP. (F) The majority of neurons are positive for GFP. (G) TH+ neurons co-express floor plate marker FOXA2. (H) TH+ neurons co-express A10 marker Calbindin. (I) TH+ neurons express A9 marker Girk2. (J) All iPSCs from the three rhesus monkeys produce teratoma tissues that represent three germ layers, including neuroectoderm, mesoderm (cartilage), and endoderm (gut epithelia), 2-3 months following injection into the SCID mice. Bar = 50 μm. Cell Reports 2013 3, 646-650DOI: (10.1016/j.celrep.2013.02.016) Copyright © 2013 The Authors Terms and Conditions

Figure S2 Host Response to Grafts, Related to Figure 2 Representative grafts in the putamen and nigra are labeled with a GFP antibody and counter stained with Nissl. The host response is revealed by staining for GFAP, CD68, CD3, CD8, HLA-DR without Nissl. Staining on spleen tissues is used as a positive control. Bar = 100 μm. Cell Reports 2013 3, 646-650DOI: (10.1016/j.celrep.2013.02.016) Copyright © 2013 The Authors Terms and Conditions