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Blood, Vol. 113, Issue 11, 2595-2604, March 12, 2009
Mesenchymal stem cell–mediated ectopic hematopoiesis alleviates aging-related phenotype in immunocompromised mice
Blood Yamaza et al.
113: 2595
Supplemental materials for: Yamaza et al
Files in this Data Supplement:
- Document 1. Supplemental methods and materials (PDF, 82.7 KB)
- Figure S1. Subcutaneous transplantation of hydroxyapatite tricalcium phosphate (HA/TCP) particles (JPG, 50.6 KB)
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Kaplan-Meier analysis of survival showed that recipient mice transplanted with subcutaneous HA/TCP particles (HA/TCP, n=12) had similar lifespan compared to age-matched control immunocompromised mice (Control, n=20).
- Figure S2. EPO treatment improved BMMSC proliferation and viability (JPG, 71.9 KB)
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(A–C) After three days rhEPO treatment (0.1 U/ml), human BMMSCs showed a significantly increased number of Brdu positive cells (A) and viable cells either with FBS (FBS+) or without FBS (FBS−) in the culture when compared to the un-treated control BMMSCs (n=3) (B). Flow cytometric analysis showed that EPO treatment induced a significantly increased expression of EPO-R in cultured human BMMSCs (n=3)(C). (D,E) Although serum EPO level was not increased in BMMSC/HA/TCP recipient mice by ELISA (D), BMMSCs derived from the recipient mice exhibited a significantly increased expression of EPO-R by Western blot analysis (E).
- Figure S3. Histological and immunohistochemical analysis of skin and spleen of hBMMSC transplant recipient mice (JPG, 89.2 KB)
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(A–C) Representative H&E staining images of skin showed that multi-cell layer of epidermis, thicker epidermis and increased number of sebaceous glands (SG) were found in 16-month-old BMMSC transplant recipient mice (Transplant, n=3) compared to the age-matched control group (Control, n=3). Collagen fibers (arrows in A) in dermis were thicker in the transplant group than that in controls. Original magnification: ×200. (D) Increased number of PCNA-positive cells was found in the epithelial cells of the epidermis (right upper panel, white open triangle arrowhead), the acinar cells of the sebaceous glands (lower panel, white open arrowheads) and the epithelial cells of the duct (lower panel, black arrow) in the recipients (Transplant) compared to the controls (Control, left panels). Original Magnification: ×200. (E) Total PCNA-positive epidermis cells were significantly higher in the recipients (Transplant, n=3) than that in the controls (Control, n=3). Bars represented standard deviation. (F) Immunohistochemical staining showed increased PCNA-positive cells in follicles of spleen from the recipients (Transplant, n=3) compared to the controls (Control, n=3). Bars represented standard deviation. (G) Immunohistochemical staining showed that B220-positive B-lymphocytes (dot circles) in spleen were more abundant in recipients (Transplant, n=3) compared to controls (Control, n=3). Original magnification: ×200.
- Figure S4. Histological and immunohistochemical analysis of brain and liver of hBMMSC transplant recipient mice (JPG, 101 KB)
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(A) TUNEL staining showed that 16-month-old BMMSC transplant recipients (Transplant) had less TUNEL-positive nuclei (arrows) in the hippocampus than the age-matched control hipocampus (Control). Original magnification: ×400. (B) H&E staining showed that the number of Purkinje cells (arrows) in the cerebellum was significantly increased in recipient mice (Transplant, n=3) as compared to controls (Control, n=3). The cellular density (open arrows) in the medulla of the recipient mice was higher than the controls (Control). Original magnification: ×200. Bars represented standard deviation. (C) Immunohistochemical staining showed that the recipients (Transplant, n=3) had significantly higher number of PCNA-positive ependimal cells of the choroid plexus (arrows) than the controls (Control, n=3). Original magnification: ×200. Bars represented standard deviation. (D) H&E staining showed degenerative hepatocytes (arrows) near the central vein (CV) in control mice (Control), but absent in age-matched BMMSC transplant recipient. Original magnification: ×200. (E) ATPase was highly expressed in hepatocytes of the transplant recipients (Transplant, n=3) compared to the controls (Control, n=3). Bars represented standard deviation. (F) However, immunohistochemical showed no significant difference of mitchondria (Mt) expression in hepatocytes between recipient mice (Transplant, n=3) and controls (Control, n=3). Bars represented standard deviation.
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