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培养的小鼠胚胎腭上皮片:使用溴脱氧尿苷对增殖活性的免疫细胞化学研究

Mouse embryonic palatal epithelial sheets in culture: an immunocytochemical study of proliferative activity using bromodeoxyuridine.

作者信息

Carette M J, Ferguson M W

机构信息

Department of Cell and Structural Biology, University of Manchester, UK.

出版信息

Epithelial Cell Biol. 1992 Jul;1(3):119-27.

PMID:1307944
Abstract

Mouse secondary palate morphogenesis is accompanied by distinctive patterns of proliferation in the palatal epithelium which latterly reflect its region-specific differentiation into oral, nasal and medial edge phenotypes. Isolated intact embryonic palatal epithelial sheets were cultured prior to, and during, the critical period of epithelial differentiation in chemically defined culture medium with, and without, 10% donor calf serum. The spatial and temporal patterns of proliferative activity were investigated by immunocytochemistry in 'pulse' and 'continuous' labelling experiments using bromodeoxyuridine (BrdUrd). Continuously labelled cultures exhibited extensive proliferation throughout the oral, nasal and medial edge regions. Pulse labelled cultures demonstrated a shift in mitotic activity from nasal to oral epithelial cells probably representing the cell turnover associated with the respective differentiated phenotypes. Medial edge cells became post-mitotic within the first 19 h of culture. Our defined culture system coupled with the immunocytochemical detection of cell proliferation using BrdUrd offers a rapid and precise method for the further investigation of palatal epithelial proliferation and its regulation by extrinsic factors.

摘要

小鼠次生腭形态发生伴随着腭上皮中独特的增殖模式,这种模式随后反映了其向口腔、鼻腔和内侧边缘表型的区域特异性分化。在化学限定培养基中,添加和不添加10%供体小牛血清的情况下,在关键上皮分化期之前和期间培养分离的完整胚胎腭上皮片。在使用溴脱氧尿苷(BrdUrd)的“脉冲”和“连续”标记实验中,通过免疫细胞化学研究增殖活性的时空模式。连续标记的培养物在整个口腔、鼻腔和内侧边缘区域表现出广泛的增殖。脉冲标记的培养物显示有丝分裂活性从鼻腔上皮细胞转移到口腔上皮细胞,这可能代表了与各自分化表型相关的细胞更新。内侧边缘细胞在培养的最初19小时内进入有丝分裂后期。我们定义的培养系统与使用BrdUrd进行细胞增殖的免疫细胞化学检测相结合,为进一步研究腭上皮增殖及其受外在因素的调控提供了一种快速而精确的方法。

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