Chen et al. Stem
-the DegludEc cardioVascular OuTcomEs trial (DEVOTE) trial) [63].
Chen et al. Stem Cell Analysis Therapy 2014, 5:36 http://stemcellres/content/5/2/RESEARCHOpen AccessThrombin promotes fibronectin secretion by bone marrow mesenchymal stem cells by way of the protease-activated receptor mediated signalling pathwaysJin Chen1,2, Yujie Ma1, Zi Wang2,three, Hengxiang Wang3, Lisheng Wang2, Fengjun Xiao2, Hua Wang2, Jianming Tan1* and Zikuan Guo2*AbstractIntroduction: Fibronectin (FN) is generally used in the improvement of serum-free media for the expansion of mesenchymal stem cells (MSCs). This study was aimed to observe if thrombin could stimulate FN secretion by human bone marrow MSCs and investigate the potential underlying mechanisms. Strategies: PCR was performed to detect the expression of the protease-activated receptors (PARs) in MSCs. Soon after thrombin treatment, the expression level and secretion of FN were observed by RT-PCR, immunofluorescence staining and ELISA, respectively, plus the activation of ERK1/2 and NF kappa B pathways was revealed by Western blotting, with or with no pre-treatment of small-molecule blockers particular for PAR-1 and ?.Formula of 1809395-84-3 The phenotypic and functional activities of thrombin-treated MSCs had been also observed.1240597-30-1 site Outcomes: PCR evaluation showed that human bone marrow MSCs expressed two subtypes of PARs, PAR-1 and PAR-2.PMID:23255394 Thrombin therapy enhanced MSCs to express FN at mRNA and protein levels and promoted FN secretion by MSCs, accompanied by potent adherence to the culture plastic. Thrombin induced prompt phosphorylation of ERK 1/2 and NF kappa B p65 plus the stimulatory effects of thrombin on FN secretion had been blunted by precise inhibitors of those signaling molecules. Blockage to PAR-1 and PAR-2 partially abrogated thrombin-elicited FN secretion by MSCs and ERK 1/2 phosphorylation, whereas that of NF kappa B p65 was unaffected. Additionally, thrombin-treated MSCs maintained the phenotypic capabilities, in vitro osteogenesis and adipogenesis capacities, and inhibitory activity on Phytohemagglutinin-induced allogeneic lymphocyte proliferation. Conclusions: Thrombin could promote FN secretion by MSCs through PAR-mediated ERK 1/2 activation, although NF kappa B might be also involved in an undefined manner.Introduction Mesenchymal stem cells (MSCs) are adult multipotent cells that have been initially isolated from bone marrow [1] and characterized by the fibroblast-like look in culture plus the capacities to kind bone, adipose and cartilage. Because the initially reported clinical trial in 1995,* Correspondence: doctortjm@yahoo; [email protected] Equal contributors 1 Fujian Provincial Essential Laboratory of Transplant Biology, Fuzhou Common Hospital, Xiamen University, Fuzhou 350025, China 2 Department of Experimental Hematology, Institute of Radiation Medicine, Beijing 100850, China Complete list of author information is offered at the end of your articleMSCs have already been increasingly used for clinical investigation ranging from immunological intervention to tissue engineering and trauma repair [2-6]. However, the quantity of MSCs is very low in bone marrow (about 0.001 to 0.01 on the mononuclear cells) and in vitro expansion is the prerequisite for their clinical application. MSCs might be easily expanded in culture media containing fetal bovine serum (FBS) from selected lots. Even so, MSCs cultured with this protocol might expose the recipients for the potentially contaminated pathogens in FBS and also the risk of sensitization elicited by xenogeneic proteins engulfed in to the c.
