SEPARATION OF THE EPIDERMAL SHEET BY DISPASE
SEPARATION OF THE EPIDERMAL SHEET BY DISPASE
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DOI：
10.1111/j.1365-2133.1983.tb01056.x
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发表时间：
1983-01-01
影响因子：
10.3
通讯作者：
OKADA, N
中科院分区：
文献类型：
--
作者：
KITANO, Y;OKADA, N
分散酶是一种细菌中性蛋白酶，它是从多粘芽孢杆菌的培养滤液中获得的。用500和1000 U/ml分散酶处理人皮肤24小时后，表皮片很容易从真皮上剥离，其下表面保留网脊。电镜观察表明，基表面由细胞组成，细胞内有许多细长的绒毛和胞质突起。虽然棘层和基底层的细胞内空间很宽，但所有桥粒及其伴随的张力丝都是完整的。通过将分散酶分离的表皮片在胰蛋白酶溶液中孵育15分钟获得表皮细胞悬浮液，并且如通过台盼蓝染料排除试验所测量的，细胞的存活率大于95%。通过以常规方式接种解离的细胞来建立角质形成细胞的培养物。
Dispase is a bacterial neutral protease which is obtained from the culture filtrate of Bacillus polymyxa. After 24-h treatment of the human skin with 500 and 1000 U/ml dispase, the epidermal sheet was easily peeled from the dermis, and its undersurface retained rete ridges. EM observation showed that the basal surface was composed of cells with numerous slender villi and cytoplasmic projections. Although the intracellular spaces of the spinous as well as the basal layer were wide, all desmosomes were intact with their accompanying tonofilaments. An epidermal cell suspension was obtained by incubating the dispase-separated epidermal sheet in trypsin solution for 15 min and the viability of the cells was more than 95% as measured by the trypan blue dye exclusion test. A culture of keratinocytes was established by seeding the dissociated cells in the usual way.