Sheets of embryonic epidermis can be isolated and cultured with or without the basal lamina.
Sheets of embryonic epidermis can be isolated and cultured with or without the basal lamina.
复制标题
可以分离胚胎表皮片并在有或没有基底层的情况下进行培养。
DOI：
10.1002/ar.1092320303
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Svoboda,KK
中科院分区：
文献类型：
--
作者：
Reenstra,WR;Svoboda,KK
在目前的研究中，我们确定了分散酶II，中性蛋白酶的浓度，和孵化时间所需的胚胎禽表皮分离。表皮可以与真皮一致地分离，具有或不具有基底层。由于Dispase II的作用具有选择性且易于控制，因此本研究表明，Dispase可作为研究表皮-细胞外基质相互作用的有力工具。在进一步的实验中，我们研究了分离的表皮片与无其基底层细胞外基质的反应。当表皮被分离而没有基膜时，基底表面泡和基底细胞质中的肌动蛋白被破坏。分离的表皮没有基底层，并在细胞外基质上培养，重组基底肌动蛋白垫并收缩水泡。然而，表皮分离无基底层和细胞外基质的情况下培养继续有水泡和一个混乱的肌动蛋白皮质垫。与此相反，与基底层分离的对照表皮在有或没有细胞外基质的情况下培养后保留平坦的基底表面和有组织的肌动蛋白皮质垫。细胞外基质已被许多研究者证明在细胞骨架组织、代谢和分化中起重要作用。细胞外基质如何相互作用和影响这些功能尚未完全阐明。使用Dispase II分离具有和不具有其基底层的组织的能力将有助于解决这些和其他上皮细胞生物学问题。
In the current study, we determine the concentration of Dispase II, a neutral protease, and the incubation time necessary for isolation of embryonic avian epidermis. The epidermis can be isolated from the dermis consistently with or without the basal lamina. Because the action of Dispase II is selective and easily controlled, this study suggests that Dispase can serve as a powerful tool for studying epidermal‐extracellular matrix interactions. In further experiments we study the response of sheets of epidermis isolated with and without its basal lamina to extracellular matrix. When the epidermis is isolated without the basal lamina, the basal surface blebs and the actin in the basal cytoplasm are disrupted. Epidermis isolated without the basal lamina and cultured on extracellular matrix reorganizes the basal actin mat and retracts the blebs. However, epidermis isolated without the basal lamina and cultured in the absence of extracellular matrix continues to have blebs and a disorganized actin cortical mat. In contrast, the control epidermis isolated with the basal lamina retains a flat basal surface and organized actin cortical mat after culturing either with or without extracellular matrix. The extracellular matrix has been shown by many investigators to play an important role in cytoskeletal organization, metabolism, and differentiation. How the extracellular matrix interacts and influences these functions has not been completely elucidated. The ability to isolate tissue consistently with and without its basal lamina using Dispase II will facilitate addressing these and other epithelial cell biology questions.