Tenchini M L, Morra F, Soranzo C, Malcovati M
Dipartimento di Biologia e Genetica per le Scienze mediche, Università di Milano, Italy.
Epithelial Cell Biol. 1995;4(4):143-55.
We previously demonstrated that spreading and clustering of in vitro cultured human keratinocytes are autocrine-induced phenomena, mediated by keratinocyte-secreted soluble factor. In this paper, the effects of this factor on spreading, number of dendrites and cell-cell contacts of the two cellular components of skin, melanocytes and fibroblasts have been studied 24 h after plating cells on uncoated plastic surfaces in MCDB153 serum-free medium or in the same medium conditioned by keratinocytes (KCM). Spreading of melanocytes present in the epidermal cell population remained constant at increasing cell density, while that of keratinocytes showed a statistically significant increase. Moreover, time-course experiments showed that the rate of spreading was faster for melanocytes. At increasing epidermal cell density, a statistically significant increase in number of dendrites and cell-cell contacts of melanocytes was observed. Similar results were obtained when melanocytes were plated both in coculture with keratinocytes (as epidermal cell cultures) or as a pure cell population in keratinocyte conditioned medium (KCM), suggesting that the observed phenomena are due to keratinocyte-secreted soluble factors and not to direct keratinocyte-melanocyte interactions. The addition of nerve growth factor (NGF) to fresh medium or addition of an inactivating anti-NGF monoclonal antibody (alpha D11) to KCM did not affect the number of dendrites or cell-cell contacts of melanocytes. Keratinocyte-secreted soluble factor(s) present in KCM also dramatically influenced morphology and cell-cell contacts of human dermal fibroblasts.
我们之前证明，体外培养的人角质形成细胞的铺展和聚集是自分泌诱导现象，由角质形成细胞分泌的可溶性因子介导。在本文中，在无血清的MCDB153培养基或角质形成细胞条件培养基（KCM）中，将细胞接种在未包被的塑料表面24小时后，研究了该因子对皮肤的两种细胞成分黑素细胞和成纤维细胞的铺展、树突数量和细胞间接触的影响。表皮细胞群体中存在的黑素细胞的铺展在细胞密度增加时保持恒定，而角质形成细胞的铺展则显示出统计学上的显著增加。此外，时间进程实验表明黑素细胞的铺展速度更快。在表皮细胞密度增加时，观察到黑素细胞的树突数量和细胞间接触有统计学上的显著增加。当黑素细胞与角质形成细胞共培养（作为表皮细胞培养物）或作为角质形成细胞条件培养基（KCM）中的纯细胞群体接种时，也获得了类似的结果，这表明观察到的现象是由于角质形成细胞分泌的可溶性因子，而不是由于角质形成细胞与黑素细胞的直接相互作用。向新鲜培养基中添加神经生长因子（NGF）或向KCM中添加失活的抗NGF单克隆抗体（αD11）均不影响黑素细胞的树突数量或细胞间接触。KCM中存在的角质形成细胞分泌的可溶性因子也显著影响人真皮成纤维细胞的形态和细胞间接触。
