Age-related changes in rat bone-marrow mesenchymal stem cell plasticity
1 Institute of Molecular Biophysics, Florida State University, Tallahassee, USA
2 Department of Biological Science, Florida State University, Tallahassee, USA
BMC Cell Biology 2011, 12:44 doi:10.1186/1471-2121-12-44Published: 12 October 2011
Additional file 1:
MSC morphology and population size in young and old bone marrow: (A) Representative phase-contrast micrographs of resulting primary isolation BM-MSCs derived prior to the first passage at 24 hours (24 h), 48 hours (48 h), day 5 and day 7. (B) Passage 0 BM-MSCs from individual young and old rats were trypsinized, and analyzed with a Cedex HiRes non-flow imaging cytometer to determine the average cell diameter of each individual MSC population. Representative phase-contrast micrographs from each experimental group are shown.
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Additional file 2:
(A) Columns show fluorescence signal intensity based on three independent differentiation experiments (n = 3) per animal. The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from younger donors (p < 0.05). Error bars designate means ± SEM (n = 4).
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Additional file 3:
(A) VEGF, EGF, IGF and G-CSF expression measured by qPCR. (B) Nkx2.5, GATA 4, cTnC, cTnI, cTnT and cTrpm expression measured by qPCR following induction with growth factors and conditioned media. Results are normalized for expression in unstimulated cells at the same time of differentiation. Data represent mean ± SEM of three biologically independent experiments realized in duplicate. The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from younger donors (p < 0.05).
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Additional file 4:
Primer sequences for specific genes Additional Statistical Analysis Fluorescence Signal Intensity Measurements. Cardiomyogenic, osteogenic, chondrogenic and adipogenic differentiation was carried out on each individual cell line from each animal (n = 4, per group). Three independent differentiation experiments (n = 3) were carried out for each cell line. Zeiss LSM Image Browser software, Rel. 4.2 was used to determine percent fluorescence intensity. Differences between groups following immunocytochemical evaluation were compared by using 1-way ANOVA. Tukey's multiple comparison test was used to establish statistical significance between experimental groups at the p < .05 (*) level. Negative controls for all immunocytochemistry experiments represent secondary antibody staining only.
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