Sato M, Ando N, Ozawa S, Miki H, Kitajima M
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
ASAIO J. 1994 Jul-Sep;40(3):M389-92. doi: 10.1097/00002480-199407000-00028.
Previously, the authors successfully grafted collagen sheets covered with cultured human esophageal epithelial cells onto latissimus dorsi muscles of athymic mice. To make a tubular artificial esophagus they developed a polyglycolic acid mesh-collagen complex tube whose inner side was epithelialized in vitro by cultured human esophageal epithelial cells. Human esophageal epithelial cells were isolated from normal mucosa resected from specimens of esophageal cancer patients and cultured on the surface of the collagen gel in which polyglycolic acid mesh was embedded. After the culture reached confluence, polyglycolic acid mesh was sutured to create a tubular form lined with cultured cell layers (restructured mucosal tube). Such tubes were wrapped in the latissimus dorsi muscle flaps of athymic rats using a fibrin glue. The animals were killed 4, 8, 20, and 28 days after grafting and the grafted tubes were studied histologically. Up to 28 days after grafting, the luminal structure was maintained without structure. Eight days after grafting, rat fibroblasts infiltrated from the muscle and neovascularization appeared in the collagen layer. The grafted epithelium remained healthy and grew to 15 cell layers by 20 days, much like normal human esophageal epithelium. Staining by anti-laminin monoclonal antibody confirmed a basement membrane.
此前，作者成功地将覆盖有培养的人食管上皮细胞的胶原片移植到无胸腺小鼠的背阔肌上。为制作管状人工食管，他们研制了一种聚乙醇酸网-胶原复合管，其内侧在体外经培养的人食管上皮细胞上皮化。人食管上皮细胞从食管癌患者标本切除的正常黏膜中分离出来，培养于包埋有聚乙醇酸网的胶原凝胶表面。培养达到汇合后，缝合聚乙醇酸网以形成内衬培养细胞层的管状结构（重组黏膜管）。将此类管子用纤维蛋白胶包裹在无胸腺大鼠的背阔肌皮瓣中。在移植后4天、8天、20天和28天处死动物，并对移植的管子进行组织学研究。移植后长达28天，管腔结构保持完整。移植后8天，大鼠成纤维细胞从肌肉浸润，胶原层出现新生血管形成。移植的上皮保持健康，到20天时生长至15层细胞，很像正常的人食管上皮。抗层粘连蛋白单克隆抗体染色证实有基底膜。
