Investigation of interstitial cells of Cajal and pacemaker mechanism of the gastrointestinal tract using new culture system

2003年，我们提出了从小鼠小肠肌层分离的细胞团的培养体系，并证实了该细胞团在体外保持了其原有的特征。用此方法分离Cajal间质细胞(ICC)，免疫大鼠，制备抗ICC的单抗。我们建立了产生抗体(AIC)的克隆。该抗体适用于形态学研究，因为多聚甲醛固定的免疫反应性稳定，我们申请了专利。我们计划鉴定与AIC结合的表位。Nakayama博士还利用该培养体系对起搏机制进行了研究。2004年，我们利用小鼠ES细胞对另一种培养体系进行了研究，并分析了肠样结构的形成。我们发现，肠样结构的形成过程模拟了肠道器官发生的过程，几个对内胚层和肠道器官发生至关重要的转录因子的表达方式与小鼠胚胎相似。我们计划利用这种培养系统来研究平滑肌和ICC的分化。

In the period of 2003We advanced the culture system of cell clusters isolated from the muscle layer of the mouse small intestine and confirmed that the cell cluster kept its original characteristic features in vitro. Using this method we isolated interstitial cells of Cajal(ICC) and immunize to rats to make a monoclonal antibody. We established the clone to produce antibody(AIC). This antibody is suitable for morphological studies because immunoreactivity was stable for paraformaldehyde fixation, and we applied for a patent. We plan to identify the epitope binding to AIC. Dr.Nakayama also advanced the study of pacemaker mechanism using this culture system.In the period of 2004The other culture system using mouse ES cells was studied and analyzed the formation of gut-like structures. We showed that formation process of gut-like structures mimicked the gut organogenesis, and several transcription factors crucial for the development of endoderm and gut organogenesis were expressed as a similar manner to mouse embryo. We plan to study differentiation of smooth muscles and ICC using this culture system.