Mouse Embryonic Stem Cells Give Rise to Gut-Like Morphogenesis, Including Intestinal Stem Cells, in the Embryoid Body Model
Mouse Embryonic Stem Cells Give Rise to Gut-Like Morphogenesis, Including Intestinal Stem Cells, in the Embryoid Body Model
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DOI：
10.1089/scd.2008.0045
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发表时间：
2009-01-01
影响因子：
4
通讯作者：
Mugishima, Hideo
中科院分区：
文献类型：
--
作者：
Konuma, Noriyoshi;Wakabayashi, Kumiko;Mugishima, Hideo
胚胎干细胞被认为是肠衰竭患者细胞替代治疗的候选细胞，因为这些细胞可以无限扩增而不会失去其多潜能表型。我们研究了小鼠ES细胞向肠样结构(包括肠干细胞)的分化能力，并确定了有效诱导形成这些结构的培养条件。在分化培养的第21天，ES细胞衍生的肠样结构(ES-Gut)在发育中的类胚体(EBS)中被重复诱导。ES肠由内胚层上皮、平滑肌层、Cajal间质细胞和肠神经元组成，呈自发性收缩。将胚胎干细胞移植到免疫缺陷小鼠的肾被膜下，可在移植物内形成由吸收细胞和杯状细胞组成的高分化上皮。移植物中有1.9%的柱状上皮细胞呈MSI-1免疫反应阳性。与含15%血清的标准ES培养相比，含0.1%二甲基亚砜的ES培养可增加EBS中ES的数量，并可增加ES与EBS的面积比。SR培养还促进上皮成熟，形成单层柱状上皮细胞，包括吸收细胞和杯状细胞。在SR条件下培养的EBS，MSI-1mRNA和蛋白的表达明显增强。综上所述，SR条件能有效地诱导胚胎干细胞的形成，并促进包括肠干细胞在内的上皮细胞的分化。这些结果表明，基于ES细胞培养系统的细胞治疗肠衰竭是可行的。
Embryonic stem (ES) cells have been proposed as candidates for cell replacement therapy in patients with intestinal failure because these cells can be expanded indefinitely without losing their pluripotent phenotype. We investigated the differentiation capacity of mouse ES cells into gut-like structures, including intestinal stem cells, and defined culture conditions for efficient induction of formation of these structures. ES cell-derived gut-like structures (ES-guts) were reproducibly induced in developing embryoid bodies (EBs) by day 21 of differentiation culture. ES-guts contained an endodermal epithelium, a smooth muscle layer, interstitial cells of Cajal, and enteric neurons and showed spontaneous contraction. Transplantation of ES-guts under the kidney capsules of immunodeficient mice induced formation of highly differentiated epithelium composed of absorptive cells and goblet cells in the grafts. Immunoreactivity for Musashi-1 (Msi-1), a marker of intestinal stem cells, was detected in 1.9% of the columnar epithelial cells in the graft. Culture with 0.1% dimethyl sulfoxide increased the numbers of ES-guts in EBs, and serum-replacement (SR) culture, in comparison to standard ES culture containing 15% serum, increased the area ratio of ES-guts to EBs. SR culture also promoted maturation of epithelium to form a single layer of columnar epithelial cells, including absorptive cells and goblet cells. Expression of Msi-1 mRNA and protein was significantly enhanced when EBs were cultured under SR conditions. In conclusion, SR conditions efficiently induce formation of ES-guts and promote differentiation of epithelium, including intestinal stem cells. These results suggest the feasibility of cell-based therapy for intestinal failure based on ES cell culture systems.