Pintucci G, Manzionna M M, Maida I, Boffi M, Boffoli D, Gallone A, Cicero R
Istituto di Biologia Generale, Faculty of Medicine and Surgery, University of Bari, Italy.
In Vitro Cell Dev Biol. 1990 Jul;26(7):659-64. doi: 10.1007/BF02624421.
A simple method to isolate and culture liver pigment cells from Rana esculenta L. is described which utilizes a pronase digestion of perfused liver, followed by sedimentation on a Ficoll gradient. A first characterization of isolated and cultured cells is also reported. They show both positivity for nonspecific esterases, and phagocytosis ability, like the cells of phagocytic lineage. Furthermore, after stimulation with a phorbol ester, these cells generate superoxide anions. At phase contrast microscope, liver pigment cells present variability in size, morphology, and in their content of dark-brown granules. Inasmuch as a cell extract obtained from cultured cells exhibits a specific protein band with dopa-oxidase activity, when run on nondenaturing polyacrylamide gel electrophoresis, liver pigment cells from Rana esculenta L. should not be considered as melanophages, but as cells that can actively synthesize melanin. The method presented here seems to be useful to more directly investigate this extra-cutaneous melanin-containing cell system and to clarify its physiologic relevance.
描述了一种从食用蛙分离和培养肝脏色素细胞的简单方法，该方法利用灌注肝脏的链霉蛋白酶消化，然后在菲可梯度上沉降。还报告了分离和培养细胞的首次特征描述。它们显示出对非特异性酯酶呈阳性，并且具有吞噬能力，类似于吞噬细胞系的细胞。此外，在用佛波酯刺激后，这些细胞会产生超氧阴离子。在相差显微镜下，肝脏色素细胞在大小、形态以及深棕色颗粒含量方面存在差异。由于从培养细胞获得的细胞提取物在非变性聚丙烯酰胺凝胶电泳上运行时显示出具有多巴氧化酶活性的特定蛋白带，因此食用蛙的肝脏色素细胞不应被视为噬黑素细胞，而应被视为能够主动合成黑色素的细胞。这里介绍的方法似乎有助于更直接地研究这个含黑色素的皮肤外细胞系统，并阐明其生理相关性。
