Mechanism of action of factors involved in regulating the cellular interaction in melanocyte

小鼠表皮成黑素细胞优先增殖的解聚表皮细胞悬浮液来自新生小鼠皮肤中的无血清成黑素细胞定义的培养基（MDM）。14天后，几乎所有主要存在于原代培养早期阶段的角质形成细胞死亡，并获得成黑素细胞的纯培养物。表皮成黑素细胞在MDMDF中显著增加，MDMDF由补充有二丁酰腺苷3 '：5'-环一磷酸（DBcAMP）和碱性成纤维细胞生长因子（bFGF）的MDM组成。在由补充有DBcAMP的MDM组成的MDMD中，表皮黑素细胞的数量增加。另一方面，表皮黑素细胞在由补充有□-黑素细胞刺激激素（MSH）的MDM组成的MDMM中被诱导分化。纯培养的原代成黑素细胞或黑素细胞在MDMDF或MDMD中进一步与补充有内皮素（ET）-1的MDMDF或MDMD从14天起培养。成黑素细胞数量的急剧增加 ...更多信息 或黑素细胞。然而，在没有ET-1的情况下，未观察到成黑素细胞或黑素细胞的数量增加。成黑素细胞或黑素细胞数量的增加与MDMDF或MDMD中与次级角质形成细胞共培养的成黑素细胞或黑素细胞的数量相当。此外，MDM中纯培养的原代成黑素细胞从14天起用补充有ET-1的MDMM进一步培养。7天后观察到成黑素细胞-黑素细胞群体中黑素细胞百分比的显著增加。然而，在没有ET-1的情况下，没有观察到黑素细胞的百分比增加。这种增加与MDMM中与次级角质形成细胞共培养的黑素细胞的增加相当。抗ET-1抗体对原代培养的MDMDF和MDMD中黑素母细胞或黑素细胞的增殖及MDMM中黑素细胞的分化均有抑制作用。这些结果表明，ET-1是参与调节小鼠表皮黑素细胞增殖和分化的角质形成细胞衍生因子的成员之一。用同样的方法证明ET-2、ET-3、白血病抑制因子、肝细胞生长因子、干细胞因子、粒细胞-巨噬细胞集落刺激因子也是角朊细胞源性因子之一。少

Mouse epidermal melanoblasts preferentially proliferated from disaggregated epidermal cell suspensions derived from newborn mouse skin in serum-free melanoblast-defined medium (MDM). After 14 days, almost all keratinocytes that existed predominantly in the early stage of primary culture died, and pure cultures of melanoblasts were obtained. Epidermal melanoblasts dramatically increased in number in MDMDF consisting of MDM supplemented with dibutyryl adenosine 3':5'-cyclic monophosphate (DBcAMP) and basic fibroblast growth factor (bFGF). Epidermal melanocytes increased in number in MDMD consisting of MDM supplemented with DBcAMP. On the other hand, epidermal melanocytes were induced to differentiate in MDMM consisting of MDM supplemented with □-melanocyte-stimulating hormone (MSH). Pure cultured primary melanoblasts or melanocytes in MDMDF or MDMD were further cultured with MDMDF or MDMD supplemented with endothelin (ET)-1 from 14 days. A dramatic increase in the number of melanoblasts … More or melanocytes was observed after 7 days. However, no increase in the number of melanoblasts or melanocytes was observed in the absence of ET-1. The increase in the number of melanoblasts or melanocytes was comparable with that of melanoblasts or melanocytes cocultured with secondary keratinocytes in MDMDF or MDMD. Also, pure cultured primary melanoblasts in MDM were further cultured with MDMM supplemented with ET-1 from 14 days. A dramatic increase in the percentage of melanocytes in the melanoblast-melanocyte population was observed after 7 days. However, no increase in the percentage of melanocytes was observed in the absence of ET-1. The increase was comparable with that of melanocytes cocultured with secondary keratinocytes in MDMM. Moreover, anti- ET-1 antibody inhibited both the proliferation of melanoblasts or melanocytes in MDMDF or MDMD and the differentiation of melanocytes in MDMM in primary culture. These results suggest that ET-1 is one member of the keratinocyte-derived factors that are involved in regulating the proliferation and differentiation of mouse epidermal melanocytes. By using the same method it was shown that ET-2, -3, leukemia inhibitory factor, hepatocyte growth factor, stem cell factor, granulocyte-macrophage colocy-stimulating factor are also one of the keratinocyte-derived factors. Less