Studies on the formation mechanism and function of prevascular network structures within multi-layered cell-sheet constructs

利用细胞片堆叠三维共培养系统，研究了层状细胞片内血管前网络形成和移植血管前细胞片形成新血管的机制。新近发现了两个编码分泌型糖蛋白和胞内蛋白的人基因，它们在内皮细胞中表达。特别是，通过用针对后一基因的siRNA处理细胞来抑制Matrigel上的EC网络形成，表明该基因可能是血管前网络形成的调节因子之一。另一方面，胎盘生长因子（PlGF）和基质金属蛋白酶-9（MMP-9）的产生在血管前细胞片中显著增加。因此，这些可溶性蛋白可能在皮下移植后的功能性吻合中起重要作用。

Using a cell-sheet staking 3-D co-culture system, the mechanisms of prevascular network formation within layered cell-sheet and neovasvularization by engrafted prevascular cell-sheets were investigated. Two human genes encoding secreted glycoprotein and intracellular protein, expressed in endothelial cells (Ecs), were newly identified. Especially, EC network formation on Matrigel was inhibited by treating the cells with siRNA for the latter gene, indicating that this gene might be one of the regulators for prevascular network formation. On the other hand, the productions of both placenta growth factor (PIGF) and matrix metalloproteinase-9 (MMP-9) significantly increased in the prevascular cell-sheets. Therefore, these soluble proteins might play an important role in the functional anastomosis after subcutaneous transplantation.