PKC활성화에 의한 골수유래 중간엽 줄기세포의 부착능 향상

Dept. of Medical Science/석사Emerging evidence suggests that cell therapy with bone marrow-derived mesenchymal stem cells (MSCs) has beneficial effects on the injured heart. However, decreased survival and/or adhesion of MSCs under ischemic conditions limit the application of cell transplantation as a therapeutic modality. This study investigated a potential method for enhancing the adhesion ability of MSCs to improve their efficacy in the ischemic heart. Because protein kinase C (PKC) is a key factor in the regulation of cellular function, treatment of MSCs with PKC activators increased cell adhesion and spreading in a dose-dependent manner, and significantly prevented MSCs from detachment. When MSCs were treated with PKC inhibitor, adhesion of MSCs was slightly diminished, and detachment was also decreased compared to treatment with PKC activator. MSCs treated with both the PKC activator and inhibitor behaved similarly to normal controls. In the 3D matrix cardiogel, treatment with PKC activator increased the number of MSCs compared with normal controls or MSCs treated with PKC inhibitor. Expression of focal adhesion kinase and cytoskeleton-associated proteins, including paxillin, vinculin, and talin was clearly increased in PMA-treated MSCs by immunoblotting and immunocytochemistry. The effect of PMA treatment on MSCs was validated in vivo. Following injection into rat hearts, the PMA-treated MSCs exhibited significant bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) fluorescence compared with control groups. Fibrosis and infarct size were reduced in rat hearts injected with PKC activator-treated MSCs compared with hearts injected with untreated MSCs. These results indicate that PKC activator is a potential target for niche manipulation to enhance adhesion of MSCs for cardiac regeneration.ope