Additional file 4 of MGFM: a novel tool for detection of tissue and cell specific marker genes from microarray gene expression data

Description of the predicted marker genes. (126KB PDF

Additional file 5 of MGFM: a novel tool for detection of tissue and cell specific marker genes from microarray gene expression data

Primer sequences. This file includes the list of all primer sequences used by PCR. (55.7KB PDF

Additional file 3 of MGFM: a novel tool for detection of tissue and cell specific marker genes from microarray gene expression data

Gel electrophoresis images. This file includes the gel electrophoresis images (Figures S1–S11). (981KB PDF

Reduced apoptosis of hypoxically preconditioned CB-MSCs.

<p>(A) Cellular caspase-3/7 activity (n = 3) and (B) LDH-release (n = 4) after 24 h of simulated ischemia with or without preceding hypoxic preconditioning (HP). * P < 0.05, ** P < 0.001 (Student’s t-test). (C) Representative microphotographs taken by light microscopy, magnification: 10x, scale bar = 200 μm. Control shows cells not exposed to hypoxic preconditioning/simulated ischemia.</p

Protein phosphorylation and gene expression in hypoxically preconditioned CB-MSCs.

<p>(A) Phosphorylation of AKT and BAD after 24 h of simulated ischemia with or without preceding hypoxic preconditioning (HP) (n = 4). (B) Representative western blot images. (C) β-actin normalized mRNA expression of BCL-XL, BCL-2 and BAG1 prior and after 24 h of simulated ischemia with or without preceding hypoxic preconditioning (n = 6). * P < 0.05, ** P < 0.01 (Student’s t-test).</p

Growth factor and cytokine expression in hypoxically preconditioned CB-MSCs.

<p>(A) β-actin normalized mRNA expression of VEGF and EGF prior (n = 9) and after (n = 6) 24 h of simulated ischemia with or without preceding hypoxic preconditioning (HP). * P < 0.05, ** P < 0.01 (Student’s t-test). (B) VEGF protein secretion on three consecutive days of simulated ischemia with (n = 12) or without (n = 11) preceding hypoxic preconditioning. HP vs. non-HP: P < 0.05 (repeated measures ANOVA). (C) MCP-1 and IL-6 protein secretion during 24 h of simulated ischemia with (n = 12) or without (n = 11) preceding hypoxic preconditioning. ** P < 0.01 (Student’s t-test).</p

Multi-lineage differentiation of cells of clonal S462 spheres.

<p>Adherent cells (left column) and dissociated sphere cells (right columns) were cultured with growth factors inducing differentiation into cells resembling (A) Schwann cells, (B) SM/Fb and (C) neurons, which are positively stained for S100/NGFR/neurofilament (Schwann cells), SMA (SM/Fb), and MAP-2 (neurons), respectively. Insert in A illustrates S100+ Schwann cells derived from a plexiform neurofibroma culture. (D) phase contrast micrographs from differentiated cells under 3 culture conditions to generate Schwann cell, SM/Fb and neuron-like cells. Differentiated cells are indicated by arrows and non-differentiated cells by arrowheads. Bars = 20 µm. NGFR, nerve growth factor; SMA, smooth muscle actin; MAP-2, microtubule associated protein-2; PNF, plexiform neurofibroma; LC, like-cells.</p

Proliferation and self-renewal of MPNST spheres.

<p>Bromodeoxyuridine incorporation assay revealed proliferation of S462 cells in the clonal spheres (secondary spheres). The increase in cell absorbance was significant for 8, 10, and 15 days in SCM versus 3 days (p<0.01 **). Changes in the frequency of sphere formation in wells containing single cells originally from adherent S462, then from dissociated spheres plated as single cells and passaged consecutively. The increase in frequency of sphere formation with increasing passage was significant (passage 1 (primary spheres) versus passages 10 (10^th spheres) and 12 (12^th spheres), P<0.01 **, passage 2 (secondary spheres) versus passages 10 (10^th spheres) and 12^th spheres), P<0.05*).</p

MSC hypoxic preconditioning strategies for treatment of ischemic cell/tissue injury.

<p>Listed are previously published studies that investigated effects of MSC hypoxic preconditioning in <i>in vitro</i> or <i>in vivo</i> models of (simulated) ischemic cell or tissue injury. If more than one preconditioning duration was tested, the most efficient is given. BM, bone marrow; AT, adipose tissue</p><p>MSC hypoxic preconditioning strategies for treatment of ischemic cell/tissue injury.</p

Comparison of MPNST adherent and sphere cultures.

<p>(A) Parental S462 cells under standard culture conditions with serum grow adherently (passage 35). (B) Floating secondary sphere after two weeks under stem cell conditions without serum. The sphere was derived from a single cell and thus was clonal. Bars = 20 µm.</p