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。

Epithelial cell biology Pub Date : 1992-07-01
M J Carette, M W Ferguson
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引用次数: 0

摘要

小鼠腭上皮的二次形态发生伴随着独特的增殖模式,这反映了其区域特异性分化为口腔、鼻和内缘表型。分离的完整的胚胎腭上皮片在上皮分化的关键时期之前和期间,分别在含10%供体牛血清和不含10%供体牛血清的化学培养基中培养。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了增殖活性的空间和时间模式。连续标记的培养物在口腔、鼻腔和内侧边缘区域显示出广泛的增殖。脉冲标记培养表明,有丝分裂活性从鼻上皮细胞向口腔上皮细胞的转变,可能代表了与各自分化表型相关的细胞周转。内侧边缘细胞在培养的前19小时内成为有丝分裂后细胞。我们定义的培养系统与BrdUrd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部因素的调节提供了一种快速、精确的方法。
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Mouse embryonic palatal epithelial sheets in culture: an immunocytochemical study of proliferative activity using bromodeoxyuridine.

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.

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