Type IV collagen aggregates promote keratinocyte proliferation and formation of epidermal layer in human skin equivalents
Type IV collagen aggregates promote keratinocyte proliferation and formation of epidermal layer in human skin equivalents
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DOI：
10.1111/exd.13328
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发表时间：
2018-05-01
影响因子：
3.6
通讯作者：
Nishiyama, Toshio
中科院分区：
文献类型：
--
作者：
Matsuura-Hachiya, Yuko;Arai, Koji Y.;Nishiyama, Toshio
从未经酶处理的晶状体囊膜中分离出的IV型胶原在生理条件下会形成凝胶，并影响细胞的活动。在人类角质形成细胞的情况下，重组的IV型胶原凝胶在单层培养中被报道抑制了增殖。在这项研究中，我们研究了从猪晶状体囊膜中提取的IV型胶原对人体皮肤替代物(HSE)表皮形成的影响。在此培养条件下制备了IV型胶原聚集体，在单层培养和凝胶培养中均能抑制角质形成细胞的增殖。在HSE中，IV型胶原聚集体在含有人真皮成纤维细胞的收缩胶原凝胶表面重组，角质形成细胞在聚集体上培养14d。有趣的是，在IV型胶原聚集的HSE中，BrdU阳性的角质形成细胞增加，表皮层厚度约为对照培养的两倍。表皮分化标志物在表皮的上层表达，真皮-表皮交界处人基底膜成分的沉积增加。这些结果表明，IV型胶原聚集体刺激了基底层角质形成细胞的增殖，并改善了表皮层的分层。
Type IV collagen isolated from lens capsule without enzymatic treatment is known to form a gel under physiological condition and influences cellular activities. In case of human keratinocytes, the suppression of proliferation on reconstituted type IV collagen gels was reported in monolayer culture. In this study, we examined effects of type IV collagen isolated from porcine lens capsule on epidermal formation in human skin equivalents (HSEs). Type IV collagen aggregates were prepared under the culture condition and the aggregates suppressed keratinocyte proliferation in monolayer culture as well as the culture on the gels. In HSEs, type IV collagen aggregates were reconstituted on the surface of contracted collagen gels containing human dermal fibroblasts and the keratinocytes were then cultured on the aggregates for 14days. Interestingly, in HSEs with type IV collagen aggregates, the BrdU-positive keratinocytes were increased and the thickness of the epidermal layer was around twice than that of control culture. Epidermal differentiation markers were expressed in the upper layer of the epidermis and the defined deposition of human basement membrane components were increased at the dermal-epidermal junction. These results indicate that the type IV collagen aggregates stimulate the proliferation of basal keratinocytes and improve the stratification of epidermal layers in HSEs.