Cario Muriel, Taieb Alain
INSERM 1035, University of Bordeaux, Bordeaux Cedex, France.
Department of Dermatology and Pediatric Dermatology, Bordeaux University Hospitals, INSERM U 1035, University of Bordeaux, Bordeaux, France.
Methods Mol Biol. 2019;1993:33-46. doi: 10.1007/978-1-4939-9473-1_3.
Melanocytes which represent around 5% of epidermal cells are located in the basal layer. To culture melanocytes we used trypsin digestion instead of dispase to obtain a cell suspension containing only basal keratinocytes and melanocytes. Melanocytes are cells which need a great attention. Indeed they dedifferentiate easily in culture as soon as they are in pure culture. Factors secreted by contaminating keratinocytes allow melanocytes to stay dendritic but by regulating their number avoid their growth. In order to age, phototype and other individual dependent factors regulate the behavior of melanocytes in vitro. Thus, microscopic examination of melanocytes has to be performed each day to adapt conditions of culture to each primary cell culture. This is the secret to have a nice melanocyte culture.
黑素细胞约占表皮细胞的5%，位于基底层。为了培养黑素细胞，我们使用胰蛋白酶消化而非分散酶来获得仅包含基底角质形成细胞和黑素细胞的细胞悬液。黑素细胞是需要格外关注的细胞。实际上，一旦处于纯培养状态，它们在培养过程中很容易去分化。污染的角质形成细胞分泌的因子可使黑素细胞保持树突状，但通过调节其数量来抑制其生长。为了使黑素细胞老化、根据光型及其他个体相关因素来调节其体外行为。因此，必须每天对黑素细胞进行显微镜检查，以使培养条件适应每种原代细胞培养。这就是成功培养出优质黑素细胞的秘诀。
