1. Volume 13, Issue 6, Pages 1173-1184 (June 2006)
Preclinical Evaluation of Adult Stem Cell Engraftment and Toxicity in the CNS of Rhesus Macaques 
Iryna A. Isakova, Kate Baker, Jason Dufour, Dina Gaupp, Donald G. Phinney 
Molecular Therapy 
Volume 13, Issue 6, Pages (June 2006)
DOI: /j.ymthe
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

2. FIG. 1
Cell surface phenotype of rhesus macaque MSCs. Histograms generated by FACS analysis show the relative staining intensity of MSCs harvested at second passage for various cluster designation (CD) antigens characteristically expressed by hematopoietic and nonhematopoietic cell lineages. The percentage of cells within the population that express the respective antigen (gray region) compared to the isotype control (black line) is indicated for each antibody evaluated.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

3. FIG. 2
Differentiation potential of rhesus macaque MSCs in vivo. (A–D) Sections of osseous tissue generated from second-passage MSCs contain an abundance of collagen (blue) and stroma formation (purple) (A), a well-defined reticulum lining the surface of collagen-rich deposits (B), an osteocyte-like cell (red arrow) residing in a well-formed lacuna (C), and blood islands containing host-derived erythrocytes (D). (E, F) Sections of osseous tissue generated from fifth-passage MSCs contain significantly diminished levels of collagen and stroma deposition. Original magnification, 400× (C, D), 200× (B), or 100× (all others).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

4. FIG. 3
Evaluation of MSC engraftment in the CNS. (A) Total amount of male DNA detected in 3-mm coronal brain slices from each transplant recipient by real-time PCR. (B) Schematic illustrating how the brain from individual transplant recipients was segregated into 3- (gray) or 6-mm (white) coronal slices, which are numbered 1–13. The red dots indicate the approximate site of intracranial MSC injections. (C) Coronal sections illustrating the approximate locations of tissue specimens from female transplant recipients shown to contain male DNA by real-time PCR. (D–M) Frozen sections of brain tissue stained with an anti-BrdU antibody (brown) and counterstained with Gil's hematoxylin (blue). Photomicrographs show BrdU-labeled engrafted donor cells (red arrows) in the dura matter lining the cortex of forebrain (D), the white matter adjacent to the cingulate (E) and reticular (F and G) gyrus, the cortex of layer 9 (H), the polymorph layer of hippocampus (I), the ventral spinocerebellar tract (J), the white matter between the cerebellar lobules (K), and the granular layer of cerebellum (L–N). Original magnification, 400× (F, H, and M) or 200× (all others).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

5. FIG. 4
Phenotype of MSCs engrafted in the CNS. (A–D) Photomicrographs of frozen brain sections stained with antibodies against CD68 (A), GFAP (B), MAP2 (C), or NeuN (D) and counterstained with DAPI (blue). Immunoreactivity (green) was evident within white matter of forebrain (A and B) and hippocampus (C and D). (E–H) Photomicrographs showing BrdU-positive (pink) donor cells (arrows) in white matter of forebrain adjacent to the reticular gyrus (E and F) and at the margin of the granular layer in the hippocampus (G and H). 3-D projection images over the z axis (0.5-μm intervals) and x axis (adjacent images) confirm that BrdU staining (pink) is localized specifically within the nucleus (blue) of cells within forebrain (F) and hippocampus (H). (I and J) Double staining for CD68 (green) and BrdU (pink) reveals a cluster of macrophages in the white matter of forebrain (I). Higher power image from the same region (J) reveals individual cells labeled with either BrdU (arrow, pink) or CD68 (arrowhead, green). (K) Double staining reveals BrdU-positive (pink) donor cells (arrows) in the granular layer of the hippocampus that lack expression of NeuN (green). (L and M) Double staining for BrdU (pink) and MAP2 (green) shows engrafted donor cells (arrows) between the polymorph and the granular layer of the hippocampus. (N–P) 3-D projection images over the z axis (0.5-μm intervals) and x axis (adjacent images) of sections double labeled for BrdU (pink) and GFAP (green) show donor cells (arrows) within white matter of forebrain (N) and within white matter tracks between cerebellar lobules (O and P). Original magnification, 600× (G), 400× (F, H, J, and M–P), 200× (A–E, I, K, and L).
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

6. FIG. 5
Home cage activity budgets. (A, B) Plotted is the proportion of total time (%) each subject performed the various behaviors in the baseline compared to the (A) first postsurgical phase (N = 10) and (B) second postsurgical phase (N = 6). Paired statistics was performed on the data collected pre- and postsurgery so that each animal was used as its own control (*P ≤ 0.05). Analyses were separated based on end points due to changes in group sizes. Abbreviations: MS, motor stereotypy; OA, other abnormal; SD, self-directed; MO, manipulate objects; ED, eating and drinking; LO, locomotive; IA, inactive; SO, social.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions

7. FIG. 6
Upper limb performance during motor testing at two levels of difficulty. (A–D) Plotted are the mean treat retrieval times for the right (A) and left (B) hand and the fastest treat retrieval times for the right (C) and left (D) hand of all experimental animals evaluated at two levels of difficulty (straight or curved rod). Motor performance was evaluated prior to surgery (Baseline) and at the first (P/S-1) and second (P/S-2) postsurgical phases.
Molecular Therapy  , DOI: ( /j.ymthe )
Copyright © 2006 The American Society of Gene Therapy Terms and Conditions