Huang Wen-Shyan, Wei Lin-Gwei, Li Jhen-Kai, Fu Keng-Yen, Huang Tai-Chun, Hsieh Pai-Shan, Huang Nien-Chi, Dai Lien-Guo, Chang Fung-Wei, Loh Shih-Hurng, Chen Yuan-Hao, Yang Bing-Heng, Shiau Chia-Yang, Wu Gwo-Jang, Dai Niann-Tzyy
Division of Plastic and Reconstructive Surgery, Taoyuan Armed Forces General Hospital, Taoyuan.
Division of Plastic and Reconstructive Surgery, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center.
Ann Plast Surg. 2019 Jan;82(1S Suppl 1):S119-S125. doi: 10.1097/SAP.0000000000001698.
The pigment melanin is produced by melanocytes, is primarily responsible for skin color, and protects it against ultraviolet rays that can cause the destruction of genetic material within the keratinocytes. To elucidate the mechanisms of many diseases associated with melanocytes, such as melanoma and albinism, or burns with uneven pigment distribution, the disease model needs to be established first. In this study, we aimed to construct the melanocyte model from patients in a short period.Sandai virus vector containing 4 stemness genes (Oct4, Sox2, Klf4, c-Myc) was transfected into human adipose-derived stem cells to produce induced pluripotent stem cells (iPSCs). Immunofluorescence staining was used to confirm the expression of specific proteins for iPSCs, including Tra-1-60, Tra-1-81, Oct-4, Sox-2, and Nango. polymerase chain reaction results also showed that specific genes of iPSCs with the ability to cause the differentiation of cells into the 3 germ layers were expressed. In our in vivo experiments, iPSCs were subcutaneously injected into nude mice to induce teratoma formation for 2 months. The morphology of the 3 germ layers was confirmed by hematoxylin and eosin staining. Furthermore, melanocytes were purified by serial induction medium, and their presence was confirmed by flow cytometry and the expression of different markers for melanocytes.
色素黑色素由黑素细胞产生，主要负责皮肤颜色，并保护皮肤免受紫外线伤害，紫外线可导致角质形成细胞内的遗传物质被破坏。为阐明许多与黑素细胞相关的疾病机制，如黑色素瘤、白化病或色素分布不均的烧伤，首先需要建立疾病模型。在本研究中，我们旨在短期内构建来自患者的黑素细胞模型。将含有4个干性基因（Oct4、Sox2、Klf4、c-Myc）的仙台病毒载体转染到人脂肪来源干细胞中以产生诱导多能干细胞（iPSC）。免疫荧光染色用于确认iPSC特异性蛋白的表达，包括Tra-1-60、Tra-1-81、Oct-4、Sox-2和Nanog。聚合酶链反应结果还表明，具有使细胞分化为3个胚层能力的iPSC特异性基因表达。在我们的体内实验中，将iPSC皮下注射到裸鼠中诱导畸胎瘤形成2个月。通过苏木精和伊红染色确认3个胚层的形态。此外，通过系列诱导培养基纯化黑素细胞，并通过流式细胞术和黑素细胞不同标志物的表达确认其存在。
