Tissue-Engineered Small Intestine and Stomach Form from Autologous Tissue in a Preclinical Large Animal Model
Tissue-Engineered Small Intestine and Stomach Form from Autologous Tissue in a Preclinical Large Animal Model
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DOI：
10.1016/j.jss.2009.03.062
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发表时间：
2009-10-01
影响因子：
2.2
通讯作者：
Grikscheit, Tracy C.
中科院分区：
文献类型：
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作者：
Sala, Frederic G.;Kunisaki, Shaun M.;Grikscheit, Tracy C.
背景组织工程化小肠、胃、大肠、食管和胃食管（GE）连接处已成功地由同基因细胞形成，并在小动物模型中用作拯救治疗。本研究的目的是确定工程化的肠和胃是否可以在自体的临床前大动物模型中产生，并确定组织工程化的肠是否保留了完整干细胞龛的特征。从6周龄的约克郡猪切除一小段空肠或胃。产生类器官单位，主要是上皮内容物的多细胞簇，并加载到可生物降解的支架管上。然后将构建体腹膜内植入自体宿主中。7周后，收集所有种植体并进行组织学和免疫组化分析。自动工程小肠和胃形成。组织工程肠结构复制了天然肠。组织学显示组织工程化小肠粘膜由柱状上皮组成，所有分化的肠细胞类型与神经支配的粘膜肌层相邻。肠上皮下肌成纤维细胞，参与干细胞龛形成的特化细胞，进行了鉴定。此外，细胞阳性的假定肠干细胞标志物，双皮质素和钙调蛋白激酶样-1（DCAMEKL-1）的表达被确定在基地的隐窝。最后，组织工程胃还形成了由胃窦型粘膜（粘液细胞和表面小窝细胞）和肌层组成的胃。我们在猪模型中成功地产生了具有正确结构的组织工程化肠，包括完整干细胞生态位的特征。据我们所知，这是首次在动物模型中以自体方式成功产生组织工程化肠，这可能更好地模拟人类宿主和人类的预期治疗途径。(C)2009 Elsevier Inc. All rights reserved.
Background. Tissue-engineered small intestine, stomach, large intestine, esophagus, and gastroesophageal (GE) junction have been successfully formed from syngeneic cells, and employed as a rescue therapy in a small animal model. The purpose of this study is to determine if engineered intestine and stomach could be generated in an autologous, preclinical large animal model, and to identify if the tissue-engineered intestine retained features of an intact stem cell niche.Methods. A short segment of jejunum or stomach was resected from 6-wk-old Yorkshire swine. Organoid units, multicellular clusters with predominantly epithelial content, were generated and loaded onto biodegradable scaffold tubes. The constructs were then implanted intraperitoneally in the autologous host. Seven wk later, all implants were harvested and analyzed using histology and immunohistochemistry techniques.Results. Autologous engineered small intestine and stomach formed. Tissue-engineered intestinal architecture replicated that of native intestine. Histology revealed tissue-engineered small intestinal mucosa composed of a columnar epithelium with all differentiated intestinal cell types adjacent to an innervated muscularis mucosae. Intestinal subepithelial myofibroblasts, specialized cells that participate in the stem cell niche formation, were identified. Moreover, cells positive for the putative intestinal stem cell marker, doublecortin and CaM kinase-like-1 (DCAMEKL-1) expression were identified at the base of the crypts. Finally, tissue-engineered stomach also formed with antral-type mucosa (mucus cells and surface foveolar cells) and a muscularis.Conclusion. We successfully generated tissue-engineered intestine with correct architecture, including features of an intact stem cell niche, in the pig model. To our knowledge, this is the first demonstration in which tissue-engineered intestine was successfully generated in an autologous manner in an animal model, which may better emulate a human host and the intended therapeutic pathway for humans. (C) 2009 Elsevier Inc. All rights reserved.