Characterization of in vitro gutlike organ formed from mouse embryonic stem cells
Characterization of in vitro gutlike organ formed from mouse embryonic stem cells
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DOI：
10.1152/ajpcell.00392.2003
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发表时间：
2004-06-01
影响因子：
5.5
通讯作者：
Takaki, M
中科院分区：
文献类型：
--
作者：
Ishikawa, T;Nakayama, S;Takaki, M
利用胚状体（EB）培养系统，我们已经从胚胎干细胞（ES）细胞（ES肠）中获得了一个功能性的类器官簇：“肠”。有许多类型的ES细胞簇，因为ES细胞具有发育成多种细胞类型的多能能力。在通过外源因子诱导特异性分化之前，我们表征了ES肠的综合生理和形态学特性。每个ES肠具有半球形（或囊状）结构，并表现出自发收缩[平均频率：13.5 +/- 8.8周期/分钟（cpm）]。通过c-Kit免疫反应性鉴定了卡哈尔间质细胞（ICC）的密集分布，并通过电子显微镜鉴定了ICC和平滑肌细胞的特定亚细胞结构。ICC经常与邻近的平滑肌细胞形成紧密接触，偶尔与其他ICC形成缝隙连接。广泛传播的细胞内Ca 2+浓度振荡产生在ES肠从聚集的c-Kit免疫阳性细胞。首次记录到ICC的平台电位（可能是起搏电位）和电慢波。这些事件是硝苯地平不敏感的，在小鼠肠道。我们目前的研究结果表明，ICC产生的节律性起搏活动有效地传播到平滑肌细胞，并驱动ES肠的自发节律性收缩。目前的ES肠道生理和形态学特性的表征铺平了道路，使适当的模型，以调查在肠道节律性的起源。
Using an embryoid body (EB) culture system, we have made a functional organlike cluster: the "gut" from embryonic stem (ES) cells (ES gut). There are many types of ES clusters, because ES cells have a pluripotent ability to develop into a wide range of cell types. Before inducing specific differentiation by exogenously added factors, we characterized comprehensive physiological and morphological properties of ES guts. Each ES gut has a hemispherical (or cystic) structure and exhibits spontaneous contractions [mean frequency: 13.5 +/- 8.8 cycles per min (cpm)]. A dense distribution of interstitial cells of Cajal (ICC) was identified by c-Kit immunoreactivity, and specific subcellular structures of ICC and smooth muscle cells were identified with electron microscopy. ICC frequently formed close contacts with the neighboring smooth muscle cells and occasionally formed gap junctions with other ICC. Widely propagating intracellular Ca2+ concentration oscillations were generated in the ES gut from the aggregates of c-Kit immunopositive cells. Plateau potentials, possibly pacemaker potentials in ICC, and electrical slow waves were recorded for the first time. These events were nifedipine insensitive, as in the mouse gut. Our present results indicate that the rhythmic pacemaker activity generated in ICC efficiently spreads to smooth muscle cells and drives spontaneous rhythmic contractions of the ES gut. The present characterization of physiological and morphological properties of ES gut paves the way for making appropriate models to investigate the origin of rhythmicity in the gut.