Generating Rho-0 Cells Using Mesenchymal Stem Cell Lines
Fig 6
Osteogenic, adipogenic and chondrogenic differentiation in 3a6 without treatment (3a6-wt) and 3a6 Rho-0 cells.
(a) Gene expressions implicated in the non-differentiation stage: Nano-HomeoBox (Nano-g), POU Class 5 Homeobox 1(Oct 3/4) and SRY (Sex determining region)-Y-box 2 (Sox-2) and SRY (Sex determining region)-Y-box 9 (Sox-9) in 3a6-wt and 3a6 Rho-0 cells. (b). Gene expressions implicated in the osteogenic process: alkaline phosphatase-4 (ALP) and osteocalcin-1 (OC-1) in basal condition in 3a6-wt and 3a6 Rho-0 cells. (c). Fold increase in the gene levels of ALP and OC-1 during osteogenic induction for 9 days; data are expressed relative to the basal condition (basal condition equal to 1). (d). Basal gene expressions implicated in the adipogenic process: fatty acid synthase (FASN) and peroxisome proliferator-activated receptor gamma (PPAR-γ) in 3a6-wt and 3a6 Rho-0 cells. (e). Fold increase in the gene levels of FASN and PPAR-γ following 9 days under adipogenic differentiation; data are expressed relative to the basal condition (basal condition equal to 1). (f). LD540, which stains lipid droplets, was used to measure adipogenic differentiation in 3a6-wt and 3a6 Rho-0 cells. Cells were cultured in standard medium (control) and in adipogenic medium for 9 days. Cells were fixed and double-stained with LD540 (red) for lipid droplets and Hoechst 33258 (blue) for DNA. Fluorescence was visualized with Olympus BX61 fluorescence microscopy and photographed at 20X. (g). Fold increase in the gene levels of CoL-I and CoL-II in the micropellet model following 21 days under chondrogenic differentiation; data are expressed relative to the basal condition (basal condition equal to 1). All data were obtained from three independent experiments, expressed as mean ±SD and analyzed by the unpaired t-test (*, p≤0.05; ** p<0.005). Basal, without induction. Osteo (Osteogenic), Adipo (Adipogenic), Chondro (Chondrogenic) cells grown in the presence of indicated medium.