Development of fetal rat intestine in organ and monolayer culture.
Development of fetal rat intestine in organ and monolayer culture.
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DOI：
10.1083/jcb.100.5.1611
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发表时间：
1985-05
期刊：
影响因子：
--
通讯作者：
Quaroni A
中科院分区：
文献类型：
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作者：
Quaroni A
肠上皮细胞的成熟和分化，证明在胎鼠小肠段，保持一个多月的悬浮器官培养，超微结构，生化和免疫学标准。在5-7天的时间内，胎儿肠碎片演变成球形结构，覆盖着一个单一的柱状上皮细胞超微结构相似的乳绒毛细胞。结构内存在松散的间充质细胞、细胞碎片和胶原蛋白。培养6天后，杯状细胞，不存在于胎儿肠在第18天，是众多的，发育良好。还观察到肠内分泌细胞。免疫荧光研究，采用特异性单克隆抗体的绒毛和隐窝细胞在体内，和各种酶的测定，已经证明了一个水平的分化和成熟的培养上皮细胞相似，但不相同的乳肠粘膜在体内。在培养物中未观察到隐窝和隐窝细胞标志物。在培养基中添加糖皮质激素导致蔗糖酶-异麦芽糖酶的诱导，但未能促进体内断奶时肠上皮的大部分功能变化。通过肠细胞角蛋白的特异性表达，在来自器官培养物的外植体中鉴定上皮细胞。分化特异性标志物，在原代培养的上皮细胞中存在，在纯上皮细胞群的选择和传代培养后丢失。这些结果表明，需要间充质和/或细胞外基质成分在维持分化状态的上皮细胞。本文所述的胎儿肠器官培养物与传统的器官和单层培养技术相比具有显著的优势，用于研究肠上皮发育和分化中所涉及的细胞和分子相互作用。
Maturation and differentiation of intestinal epithelial cells was demonstrated in segments of fetal rat small intestine, maintained for more than a month in suspension organ culture, by ultrastructural, biochemical, and immunological criteria. Over a 5-7 d period, fragments of fetal intestine evolved into globular structures covered with a single columnar epithelium ultrastructurally similar to suckling villus cells. Loose mesenchymal cells, cellular debris, and collagen were present inside the structures. After 6 d in culture, goblet cells, not present in the fetal intestine at day 18, were numerous and well developed. Intestinal endocrine cells were also observed. Immunofluorescence studies employing monoclonal antibodies specific for villus and crypt cells in vivo, and various enzyme assays, have demonstrated a level of differentiation and maturation of the cultured epithelial cells similar but not identical to that of suckling intestinal mucosa in vivo. Crypts and crypt cell markers were not observed in the the cultures. Addition of glucocorticoids to the culture medium resulted in the induction of sucrase-isomaltase but failed to promote most of the functional changes characteristic of the intestinal epithelium at weaning in vivo. Epithelial cells were identified in explants derived from the organ cultures by their specific expression of intestinal cytokeratin. Differentiation-specific markers, present in the epithelial cells in primary cultures, were lost upon selection and subculturing of pure epithelial cell populations. These results suggest a requirement for mesenchymal and/or extracellular matrix components in the maintenance of the differentiated state of the epithelial cells. The fetal intestinal organ cultures described here present significant advantages over traditional organ and monolayer culture techniques for the study of the cellular and molecular interactions involved in the development and differentiation of the intestinal epithelium.