Lipomas are benign adipose tissue tumors of unknown etiology, which can vary in size, number, body localization and cell populations within the tissue. were at different stages of osteogenesis. Differences observed between LDSCs and ADSCs are probably due to the GSK1292263 distinct molecular signature and their commitment in the tissue that governs their different capacity and fate during adipogenic and osteogenic induction in vitro despite their similar mesenchymal phenotype. (QT00079247), (QT00073549), (QT00210840), GSK1292263 (QT00232771), (QT00998102), (QT00093128), (QT00029841), (QT00030261), and (QT00014091). The protocol conditions were: (1) enzyme activation: 3 min at 95 C (1 cycle); (2) denaturation: 3 s at 95 C and annealing/extension (with data acquisition): 30 s at 60 C (40 cycles). The specific binding of primers was confirmed by melting GSK1292263 curve analysis and specific length product visualization on electrophoresis gel. The expression level of each target gene was normalized to the glyceraldehyde-3-phosphate dehydrogenase housekeeping gene expression ( 0.05 was considered as significant. 3. Results 3.1. Analysis of Mesenchymal Stem Cell Phenotype In Figure 1 the morphology of LDSCs (a,b) and ADSCs (c,d) is presented. There were no differences in morphology between LDSCs and ADSCs either at day 3 after isolation (Figure 1a,c) or at passage 1 day 4 (Figure 1b,d). Open in a separate window Figure 1 Morphology of lipoma-derived stem cells (LDSCs) (a,b) and adipose-derived stem cells (ADSCs) (c,d) cultures; Images were acquired at day 3 after isolation (a,c) and at day 4 after passage 1 (b,d); phase contrast with objective magnification 10; cells are spindle-like in shape which is typical for mesenchymal stem cells. Flow cytometric analysis (Figure 2aCd) showed high expression of surface stem cell marker CD105 in both LDSCs (Figure 2a) and ADSCs (Figure 2c) at passage 2, just before differentiation Elf1 assays. Non-specific antibody binding was excluded by using isotype control (Figure 2b,d). Open in a separate window Figure 2 Flow cytometric analysis of CD105 cell surface marker expression in LDSCs (a) and ADSCs (c) at passage 2 (representative histograms per each group of samples with % of CD105 positive cells presented as mean SD, n (LDSCs) = 6 and n (ADSCs) = 4); corresponding isotype controls (b,d); Relative expression of (e) and (f) genes in LDSCs and ADSCs at passage 2 (day 0 in differentiation assays), normalized to and stem cell markers expression (Figure 2e,f) confirmed that both LDSCs and ADSCs express these genes at passage 2. Slightly higher expression of in ADSCs compared to LDSCs (Figure 2e) was not statistically significant (= 0.1). We also analyzed expression levels of genes characteristically expressed during osteogenesis (and and and expression was higher in LDSCs compared to ADSCs while expression was higher in ADSCs compared to LDSCs ( 0.05). Open in a separate window Figure 3 Relative expression of (a), (b), (c), (d), (e) and (f) genes in LDSCs and ADSCs at passage 2 (day 0 in differentiation assays), normalized to and expression ( 0.05); scatterplots with median; n (LDSCs) = 6 and n (ADSCs) = 4 for all genes. 3.2. Adipogenic Differentiation Adipogenic differentiation of both LDSCs and ADSCs was analyzed after 21 days of cultivation in adipogenic medium (AM). As control, cells were cultivated in standard medium (DM) under the same conditions. Characteristic adipocyte-like phenotype and the presence of lipid droplets were noticed in both LDSCs (Figure 4a,b) and ADSCs (Figure 4e,f) after 21 days of differentiation. However, lipid droplets were noticeably significantly more present in.