Open Access Research article

Phenotype, donor age and gender affect function of human bone marrow-derived mesenchymal stromal cells

Georg Siegel1, Torsten Kluba2, Ursula Hermanutz-Klein1, Karen Bieback3, Hinnak Northoff1 and Richard Schäfer14*

Author Affiliations

1 Institute of Clinical and Experimental Transfusion Medicine (IKET), University Hospital Tübingen, Otfried-Müller-Strasse 4/1, Tübingen, D-72076, Germany

2 Department of Orthopaedic Surgery, University Hospital Tübingen, Hoppe-Seyler-Strasse 3, Tübingen, D-72076, Germany

3 Institute of Transfusion Medicine and Immunology, German Red Cross Blood Service of Baden-Württemberg-Hessen, Medical Faculty Mannheim, Heidelberg University, Friedrich-Ebert Strasse 107, Mannheim, D-68167, Germany

4 Department of Neurosurgery, Stanford Institute for Neuro-Innovation and Translational Neurosciences, Stanford University School of Medicine, 1201 Welch Road, Stanford, CA, 94305-5487, USA

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BMC Medicine 2013, 11:146  doi:10.1186/1741-7015-11-146

Published: 11 June 2013

Additional files

Additional file 1: Figure S4:

Flow cytometry gating strategy. FSC–SSC gating to separate debris from intact cells (G1). Dead cells were excluded by uptake of 7-AAD (G2 on 7-AAD negative (= live) cells). Percentage analysis of antigen-positive cells and fluorescence intensity was performed with FlowJo-7.2.5 software. For compensation of unspecific antibody binding, the positivity of the respective matched isotype control was subtracted from all samples.

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Additional file 2: Figure S5:

Flow cytometry density plots. Density plots from flow cytometric analysis show a representative BM-MSC preparation (P1). Overlay density plots show gated cells after excluding non-viable cells and debris. Green: specific antibody; red: isotype control.

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Additional file 3: Table S1:

Donor distribution and data for CFU-F analyses.

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Additional file 4: Figure S1:

Expression of androgen receptor and FGFR3 by BM-MSCs. No difference was detected in androgen receptor and FGFR3 expression between BM-MSC preparations of female and male donors (A, B) and no significant correlation was found between these receptors and donor age (C, D) (n = 14 (7 female, 7 male)). Lower detection limits (ELISA): Androgen receptor: 113 pg/ml; FGFR3: 55 pg/ml. Two-tailed Student’s t-test and Spearman two-tailed correlation test, error bars: SD.

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Additional file 5: Figure S2:

Adipogenic differentiation potential of MSCs. The percentage of CD10+ cells (n = 27) and the specific antibody mediated fluorescence per cell (ΔGeo Mean) of CD119 (n = 22) correlated positively with the induction of PPARγ mRNA under adipogenic differentiation (A-D). Moreover, the percentage of GD2+ cells and the specific antibody mediated fluorescence per cell (ΔGeo Mean) of GD2 correlated negatively with the induction of LPL mRNA under adipogenic differentiation (n = 26) (E, F). Hereby, we identified two phenotypes with either higher (CD10, CD119) or lower (GD2) adipogenic differentiation potential within the BM-MSC preparations. Spearman two-tailed correlation test (*P <0.05; **P <0.01).

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Additional file 6: Figure S3:

Secretion profile of MSC trophic factors. BM-MSCs secreted the highest concentrations of VEGF-A and HGF, followed by LIF, Angiopoietin-1, bFGF and NGFB (n = 11) (A). Correlation analyses of the secreted factors to markers potentially defining MSC subpopulations revealed a significant negative correlation for HGF secretion to the expression of CD71, CD140b and Galectin 1 (n = 11 except for Galectin 1 (n = 9)) (B); no positive correlation of the tested markers to the secretion of trophic factors was identified. Neither donor age nor gender affected the secretion of trophic factors (C, D); no correlation of the marker expression to the Angiopoietin-1 “(non-)secretor” status of the MSCs was identified (n = 11) (E, F). Lower detection limits (Luminex® and ELISA): NGF-b: 3.9 pg/ml; LIF: 2.5 pg/ml; FGF-b: 13.2 pg/ml; VEGF-A: 11.2 pg/ml; HGF: 2.2 pg/ml; Angiopoietin-1: 3.45 pg/ml; BMP4: 1.04 pg/ml. ANOVA analysis of variance followed by Tukey`s Multiple Comparison Test, Two-tailed Student’s t-test and Spearman two-tailed correlation test (*P <0.05; **P <0.01; ***P <0.001). Error bars: SD.

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Additional file 7: Table S2:

Donor variations and data of Angiopoietin-1 secretion.

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