Godwin Lauren S, Castle Joanna T, Kohli Jaskaren S, Goff Philip S, Cairney Claire J, Keith W Nicol, Sviderskaya Elena V, Bennett Dorothy C
Molecular Cell Biology Group, Biomedical Sciences Research Centre, St. George's, University of London, London, United Kingdom.
Curr Protoc Cell Biol. 2014 Jun 3;63:1.8.1-20. doi: 10.1002/0471143030.cb0108s63.
Located in the basal epidermis and hair follicles, melanocytes of the integument are responsible for its coloration through production of melanin pigments. Melanin is produced in lysosomal-like organelles called melanosomes. In humans, this skin pigmentation acts as an ultraviolet radiation filter. Abnormalities in the division of melanocytes are quite common, with potentially oncogenic growth usually followed by cell senescence producing benign naevi (moles), or occasionally melanoma. Therefore, melanocytes are a useful model for studying melanoma, as well as pigmentation and organelle transport and the diseases affecting these mechanisms. This chapter focuses on the isolation, culture, and transfection of human and murine melanocytes. The first basic protocol describes the primary culture of melanocytes from human skin and the maintenance of growing cultures. The second basic protocol details the subculture and preparation of mouse keratinocyte feeder cells. The primary culture of melanocytes from mouse skin is described in the third basic protocol, and, lastly, the fourth basic protocol outlines a technique for transfecting melanocytes and melanoma cells.
位于基底表皮和毛囊中的皮肤黑素细胞通过产生黑色素色素来决定皮肤的颜色。黑色素在称为黑素小体的溶酶体样细胞器中产生。在人类中，这种皮肤色素沉着起到紫外线辐射过滤器的作用。黑素细胞分裂异常相当常见，潜在致癌性生长通常随后是细胞衰老，产生良性痣（痣），或偶尔产生黑色素瘤。因此，黑素细胞是研究黑色素瘤、色素沉着、细胞器运输以及影响这些机制的疾病的有用模型。本章重点介绍人和小鼠黑素细胞的分离、培养和转染。第一个基本方案描述了从人皮肤中进行黑素细胞的原代培养以及生长培养物的维持。第二个基本方案详细介绍了小鼠角质形成细胞饲养层细胞的传代培养和制备。第三个基本方案描述了从小鼠皮肤中进行黑素细胞的原代培养，最后，第四个基本方案概述了一种转染黑素细胞和黑色素瘤细胞的技术。
