Dermatological Research Group of the Hungarian Academy of Sciences, Szeged, Hungary.
PLoS One. 2011 Feb 23;6(2):e17197. doi: 10.1371/journal.pone.0017197.
In previous work we described a novel culture technique using a cholera toxin and PMA-free medium (Mel-mix) for obtaining pure melanocyte cultures from human adult epidermis. In Mel-mix medium the cultured melanocytes are bipolar, unpigmented and highly proliferative. Further characterization of the cultured melanocytes revealed the disappearance of c-Kit and TRP-1 and induction of nestin expression, indicating that melanocytes dedifferentiated in this in vitro culture. Cholera toxin and PMA were able to induce c-Kit and TRP-1 protein expressions in the cells, reversing dedifferentiation. TRP-1 mRNA expression was induced in dedifferentiated melanocytes by UV-B irradiated keratinocyte supernatants, however direct UV-B irradiation of the cells resulted in further decrease of TRP-1 mRNA expression. These dedifferentiated, easily accessible cultured melanocytes provide a good model for studying melanocyte differentiation and possibly transdifferentiation. Because melanocytes in Mel-mix medium can be cultured with human serum as the only supplement, this culture system is also suitable for autologous cell transplantation.
在之前的工作中，我们描述了一种新的培养技术，使用霍乱毒素和无 PMA 培养基（Mel-mix）从成人表皮中获得纯黑素细胞培养物。在 Mel-mix 培养基中，培养的黑素细胞呈双极、无色素、高度增殖。对培养的黑素细胞的进一步特征分析表明，c-Kit 和 TRP-1 的消失以及巢蛋白表达的诱导，表明黑素细胞在这种体外培养中去分化。霍乱毒素和 PMA 能够诱导细胞中 c-Kit 和 TRP-1 蛋白的表达，从而逆转去分化。UV-B 照射角质形成细胞上清液可诱导去分化黑素细胞中 TRP-1 mRNA 的表达，但细胞的直接 UV-B 照射导致 TRP-1 mRNA 表达进一步降低。这些去分化的、易于获得的培养黑素细胞为研究黑素细胞分化和可能的转分化提供了良好的模型。由于 Mel-mix 培养基中的黑素细胞可以用人血清作为唯一补充进行培养，因此该培养系统也适用于自体细胞移植。
