Cells were cultured in serum free medium for 24 h with 0. 01 umol/L of simvastatin, 0. 01 umol/L of simvas Dasatinib msds tatin plus 50 n M of wortmannin, or blank control. Cell viabi lity was evaluated using the MTT assay and flow cytometry according to the manufacturers instructions. Effect of simvastatin on the release of VEGF of bone marrow derived MSCs in vitro To examine whether simvastatin enhance the release of VEGF by bone marrow derived MSCs, a total of 1 104 MSCs were plated in serum free medium with different doses of simvastatin on 48 well plates. VEGF levels in conditioned medium were measured with VEGF ELISA kits Inhibitors,Modulators,Libraries 24 h after treatment. Statistical analysis All values were expressed as mean SD. Students unpaired t test was used to compare differences between every two groups.
Comparisons of parameters among three or four groups were made by one way ANOVA, followed by Scheffe multiple Inhibitors,Modulators,Libraries comparison test. Compari sons of the time course of the LDPI index were made by 2 way ANOVA for repeated measures, followed by Scheffe multiple comparison tests. A probability value 0. 05 was considered statistically significant. Results Identification of bone marrow derived MSCs During the primary cell culture, the attached cells stretched and took the shape of a typical spindle Inhibitors,Modulators,Libraries shaped fibroblast phenotype. These adherent cells could be readily expanded in vitro by successive cycles of trypsi nization, seeding and culture every 3 days for 15 pas sages without visible morphologic change. Flow cytometry examination showed that these cells were negative for CD34 and CD45, but positive for CD44 and CD29.
Thus, we designated these fibroblasts like Inhibitors,Modulators,Libraries cell populations as MSCs. Combination therapy increases blood perfusion To determine whether Inhibitors,Modulators,Libraries simvastatin or MSCs treatment could stimulate the blood reperfusion in ischemic limb, mice were treated with simvastatin or MSCs or vehicle, the blood reperfusion was examined at day 0, 10 and 21 after the treatment by LDPI. LDPI showed that blood flow in the ischemic hindlimb was decreased equally in all four groups http://www.selleckchem.com/products/Nilotinib.html immediately after surgery. Over the sub sequent 21 days, blood perfusion of the ischemic hin dlimb notably improved in the treatment groups The laser Doppler perfusion index was signifi cantly higher in the simvastatin group, the MSCs group and the combination group than in the control group on day 10 after treatment and showed further improve ment afterwards on day 21. The LDPI index was the highest in the combination group among the four groups. The normal value of LDPI index was 1. 00 0. 03 in this study.