향상된 혈관신생을 위한 온도감응성 하이드로젤을 이용한 혈관내피세포 평활근세포 공배양 세포패치 전달

Abstract

Cell based therapies have been chosen as an attractive source for therapeutic angiogenesis. However obtaining a stable vascular structure remains a challenge because of poor interaction of transplanted cells with native tissue and low engraftment efficiency. In this study we designed a co-cultured endothelial cells (ECs) and smooth muscle cells (SMCs) cell patches for regeneration of mature vasculatures. In vitro characterization of co-cultured cells revealed that extracellular matrix (ECM) of all groups developed more with respect of time. Furthermore, in case of ECs in co-cultured cells changed their morphology to mesh-like tuble connected with each other. Secreted growth factor showed increasing trend, allowing for the formation of extended tubular network. Confluent cell patches with ECs and SMCs were successfully fabricated on cell adhesive peptide incorporated Tetronic®-tyramine hydrogel and the detachment and transfer of cell patch was rapidly induced within 10 minutes by expansion of the hydrogels when the temperature was decreased to 4oC. The cell patch simply transferred the model surface of coverglass and distinguishable alteration in morphology of human umbilical vein endothelial cells (HUVECs) was found in cell patch from mixed group which was tube like structure. Finally, we investigated therapeutic effects of cell patch which is transplanted into hindlimb ischemic model. It retarded the rate of necrosis as compare to HUVECs and SMCs patch. By immunohistological analysis, it was confirmed that the number of arterioles and capillaries was significantly enhanced and implanted cell patch was partially involved in vessel formation. In summary, the simple transplantation of the co-cultured cell patch using a hydrogel system could assist enhanced therapeutic angiogenesis through the regeneration of matured vascular structures.