Sala Frédéric G, Kunisaki Shaun M, Ochoa Erin R, Vacanti Joseph, Grikscheit Tracy C
Developmental Biology and Regenerative Medicine Program, Saban Research Institute, Childrens Hospital Los Angeles, Los Angeles, California 90027, USA.
J Surg Res. 2009 Oct;156(2):205-12. doi: 10.1016/j.jss.2009.03.062. Epub 2009 May 3.
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.
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.
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 (DCAMKL-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.
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.
利用同基因细胞已成功构建出组织工程化小肠、胃、大肠、食管和胃食管（GE）交界处，并将其用作小动物模型的挽救疗法。本研究的目的是确定能否在自体临床前大动物模型中生成工程化肠和胃，并确定组织工程化肠是否保留完整干细胞龛的特征。
从6周龄的约克夏猪身上切除一小段空肠或胃。生成类器官单元，即主要含有上皮成分的多细胞簇，并将其加载到可生物降解的支架管上。然后将构建体腹腔内植入自体宿主。7周后，取出所有植入物，使用组织学和免疫组织化学技术进行分析。
形成了自体工程化小肠和胃。组织工程化肠结构复制了天然肠的结构。组织学显示，组织工程化小肠黏膜由柱状上皮组成，所有分化的肠细胞类型都与有神经支配的黏膜肌层相邻。鉴定出了参与干细胞龛形成的肠上皮下肌成纤维细胞，即特殊细胞。此外，在隐窝底部鉴定出了假定的肠干细胞标志物双皮质素和钙调蛋白激酶样-1（DCAMKL-1）表达阳性的细胞。最后，还形成了具有胃窦型黏膜（黏液细胞和表面小凹细胞）和肌层的组织工程化胃。
我们在猪模型中成功生成了具有正确结构的组织工程化肠，包括完整干细胞龛的特征。据我们所知，这是首次在动物模型中以自体方式成功生成组织工程化肠，这可能更好地模拟人类宿主和人类预期的治疗途径。
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