BACKGROUND: Mesenchymal stem cells (MSCs), with low immunogenicity, can regulate cellular immunity and mitigate graft rejection, which has a good prospect in tissue engineering. However, it is rarely present in bone marrow. OBJECTIVE: To explore an isolation and culture method of the rabbit bone marrow-derived MSCs, to observe the biological characteristics and differentiation potential of bone marrow-derived MSCs.METHODS: MSCs were isolated from rabbit tibia bone marrow by combination of gradient centrifugation and different adherent method, then proliferation in vitro. Morphology was examined by phase contrast microscopy, and the growth curve of cultured MSCs was drawn via MTT results. MSCs were treated with osteogenetic inductor (L-DMEM/F12, 10% fetal bovine serum, 0.1 μmol/L dexamethasone, 200 μmol/L vitamin C, 10 mmol/L β-phosphoglycerol), adipose inductor (L-DMEM/F12, 10% FBS, 1 μmol/L dexamethasone, 200 μmol/L antifani, 0.5 mmol/L IBMX, 10 μg/mL insulin), and chondrocytes inductor (L-DMEM/F12, 10% FBS, 10 μg/L TGF-β1, 0.1 μmol/L dexamethasone, 50 μmol/L vitamin C, 6.25 mg/L insulin) to differentiated into osteoblast, dipocytes and chondrocytes. And the differentiated cells were identified by alkaline phosphatase staining, oil red O staining, and toluidine blue staining, respectively.RESULTS AND CONCLUSION: Bone marrow-derived MSCs can be isolated and cultured by the combination of gradient centrifugation and different adherent method in vitro, which have the better potentiality of proliferation and multi-directional differentiation. Mostly of the primary and passaged cells were spindle-shaped. After osteogenetic induction, cells were positive to alkaline phosphatase staining. Oil red O staining showed that red lipid droplet existed in adipose cells, and toluidine blue staining showed that toluidine blue was positive after chondrocytes induction.

Objective: To investigate the effects of sulforaphane (SFN) on proliferation , migration and invasion of human renal carcinoma cells and its mechanism. Methods: The cultured human renal carcinoma cells 786⁃O were divided into control group (0 μmol/L) and SFN group (5 , 10 , 20 μmol/L) . The activated proliferation of cells was detected by CCK⁃8 ; the effect of SFN on migration of 786⁃O cells was detected by scratch healing assay and Transwell cell migration assay; the effect of SFN on the invasion ability of 786⁃O cells was detected by Transwell cell invasion ability assay; Western blot and qRT⁃PCR were used to detect the effects of SFN on the expression of epithelial⁃mesenchymal transition (EMT) Ⅳrelated proteins and mRNA. The effect of SFN on the expression of NF⁃κB signaling pathway was detected by Western blot. Results: After SFN treatment for 24 , 48 and 72 h , the proliferation activity of 786⁃O cells decreased with the increase of SFN concentration ; compared with the control group , the cell migration and invasion ability of SFN⁃treated group were significantly reduced ; with the increase of SFN concentration , the mRNA and protein expression levels of E ⁃cadherin in 786⁃O cells increased , while the mRNA and protein expression levels of N ⁃cadherin and Vimentin decreased ; the levels of NF⁃κB signaling pathway related protein phosphorylated p65 and phosphorylated IκBα decreased with the increase of SFN concentration. Conclusion SFN may inhibit the proliferation , migration and invasion of human renal carcinoma cells by regulating the EMT process of renal carcinoma through inhibition of NF⁃κB signaling pathway.