온도감응성 하이드로겔을 기반으로 배양된 근육세포시트를 이용한 신생 혈관 생성 치료기술 개발

Abstract

Peripheral arterial disease (PAD) represents altered structure and function of arteries caused by accumulated plaques. Although there have been many studies for treating these diseases by direct injection of various types of therapeutic cells, low cell engraftment efficiency and diffusion of the transplanted cells have been major problems. In the present study, we developed C2C12 myoblast layer that can be transplanted by transfer printing method using thermo-sensitive hydrogels, and investigated their efficacy in therapeutic angiogenesis. In vitro, we observed robust cell-cell contacts and stable expression of the ECM proteins in our myoblast layer. And then to confirm the therapeutic effect of the myoblast in vivo, we cultured C2C12 myoblasts on RGD incorporated thermo-sensitive hydrogel for 24 hr and transferred them to the hindlimb ischemia tissue of athymic mice directly from the hydrogel by application of 4 oC saline solution on the hydrogel. The transferred myoblast layer was retained in a longer period of time than intramuscularly injected single cells, and morphology of the mice, and Laser Doppler Perfusion (28 days) supported that the myoblast layer enhanced therapeutic effects on the ischemic tissue. In summary, transplantation of the C2C12 myoblast layer using tissue transfer printing method would represent a new approach for therapeutic angiogenesis for treating PAD.