Clark C R, Taylor J D, Tchen T T
In Vitro Cell Dev Biol. 1987 Jun;23(6):417-21. doi: 10.1007/BF02623857.
Two key modifications of the previously reported method for isolation of goldfish xanthophores allowed the isolation and establishment of primary cultures of terminally differentiated melanophores from the Black Moor goldfish (Carassius auratus). First, pretreatment with 10(-4) M epinephrine causing aggregation of the melanosomes and collapse of the dendrites, prevents damage to the melanophores during tissue dissociation and melanophore isolation. Second, maintenance of these cells in culture was successful only when the culture medium was supplemented with fish serum. The purified melanophores attached, flattened, and were maintained in culture for up to 3 mo. Although the morphology of the cultured melanophores is less dendritic than their in vivo counterparts, the melanophores translocate melanosomes in a normal manner except that they exhibit enhanced sensitivity to epinephrine. This epinephrine-induced pigment aggregation, as well as the redispersion of pigment after the removal of epinephrine, can occur in the presence of ethylene glycol-bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and absence of Ca2+.
对先前报道的金鱼黄色素细胞分离方法进行的两项关键改进，使得能够从黑 Moor 金鱼（Carassius auratus）中分离并建立终末分化黑素细胞的原代培养物。首先，用 10⁻⁴ M 肾上腺素预处理会导致黑素体聚集和树突塌陷，从而防止在组织解离和黑素细胞分离过程中对黑素细胞造成损伤。其次，只有当培养基添加鱼血清时，这些细胞才能成功维持培养。纯化后的黑素细胞附着、铺展，并在培养中维持长达 3 个月。尽管培养的黑素细胞形态比其体内对应物的树突状程度低，但黑素细胞以正常方式转运黑素体，只是它们对肾上腺素表现出增强的敏感性。这种肾上腺素诱导的色素聚集以及去除肾上腺素后色素的再分散，可在存在乙二醇双（β-氨基乙基醚）-N,N,N',N'-四乙酸且无 Ca²⁺ 的情况下发生。
