细胞角蛋白K19在体外培养人角质形成细胞中的时空序列表达。

M H Lu, P C Yang, L T Chang, C F Chao
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摘要

独特的细胞角蛋白K19在单纯上皮细胞、非角化层状鳞状上皮基底细胞、胚胎期表皮细胞和鳞状癌细胞中特异性表达,但在成人表皮中不表达。有趣的是,当表皮细胞在体外培养时,K19在基底层上重新表达。K19的表达作为表皮细胞生长和分化的标志。为了阐明培养的角质细胞中K19的时间和空间序列表达,采用两阶段人角质细胞培养系统,通过免疫印迹和免疫组织化学检测增殖和分化阶段角质细胞中K19的表达。根据我们的结果,K19在培养的人角质形成细胞中增殖期不表达,但在培养基改为分化培养基后3天在角质形成细胞中重新表达。免疫组化观察发现,K19在培养前3周持续表达于角质形成细胞的上基底层,而在基底层未检测到K19。当角化细胞用“插入培养皿”培养时,K19在由内腔和外腔培养基滋养的所有层角化细胞中持续表达。K19在两种不同培养体系中的表达差异似乎表明,K19在角质形成细胞中的表达下调与培养基供应方向有关。
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Temporal and spatial sequence expression of cytokeratin K19 in cultured human keratinocyte.

The unique cytokeratin K19 specifically expresses in simple epithelial cells, basal cells of non-keratinized stratified squamous epithelium, epidermal cells during the embryonic stage and squamous carcinoma cells, but it is not expressed in adult epidermis. Interestingly, when epidermal cells are cultured in vitro, K19 is re-expressed in the supra-basal layer. K19 expression was used as a marker for epidermal cell growth and differentiation. In order to clarify the temporal and spatial sequential expression in cultured keratinocyte, two-stage human keratinocyte culture systems were used to examine K19 expression in keratinocytes in a proliferation and differentiation stages through immunoblotting and immunohistochemistry assay. According to our results, K19 was not expressed in cultured human keratinocytes in the proliferation stage but was re-expressed in keratinocytes three days after the cultured medium was changed to a differentiation medium. Immunohistochemical observation revealed that K19 was persistently expressed in the supra-basal layer of cultured keratinocytes during first three weeks of culturing, but none was detectable in the basal cell layer. When keratinocytes were cultured with an "inserted cultured dish," K19 was persistently expressed in all layers of keratinocytes nourished by medium both from an inner chamber and an outer chamber. The different expression of K19 in these two different culture systems seemed to indicate that down regulation of K19 expression in keratinocyte was related to the direction of medium supply.

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