Tian Ting, Zhang Ru-Zhi, Yang Yu-Hua, Liu Qi, Li Di, Pan Xiao-Ru
1 The Dermal and Venereal Department, The Third Affiliated Hospital of Suzhou University , Changzhou, China .
2 The Dermal and Venereal Department, Bengbu Medical College , Bengbu, China .
Cell Reprogram. 2017 Apr;19(2):116-122. doi: 10.1089/cell.2016.0032. Epub 2017 Feb 7.
The objective of the authors has been to obtain multilineage-differentiating stress-enduring cells (Muse cells) from primary cultures of dermal fibroblasts, identify their pluripotency, and detect their ability to differentiate into melanocytes. The distribution of SSEA-3-positive cells in human scalp skin was assessed by immunohistochemistry, and the distribution of Oct4, Sox2, Nanog, and SSEA-3-positive cells was determined by immunofluorescence staining. The expression levels of Sox2, Oct4, hKlf4, and Nanog mRNAs and proteins in Muse cells were determined by reverse transcription polymerase chain reaction (RT-PCR) analyses and Western blots, respectively. These Muse cells differentiated into melanocytes in differentiation medium. The SSEA-3-positive cells were scattered in the basement membrane zone and the dermis, with comparatively more in the sebaceous glands, vascular and sweat glands, as well as the outer root sheath of hair follicles, the dermal papillae, and the hair bulbs. Muse cells, which have the ability to self-renew, were obtained from scalp dermal fibroblasts by flow cytometry sorting with an anti-SSEA-3 antibody. The results of RT-PCR, Western blot, and immunofluorescence staining showed that the expression levels of Oct4, Nanog, Sox2, and Klf4 mRNAs and proteins in Muse cells were significantly different from their parental dermal fibroblasts. Muse cells differentiated into melanocytes when cultured in melanocyte differentiation medium, and the Muse cell-derived melanocytes expressed the melanocyte-specific marker HMB45. Muse cells could be obtained by flow cytometry from primary cultures of scalp dermal fibroblasts, which possessed the ability of pluripotency and self-renewal, and could differentiate into melanocytes in vitro.
作者的目标是从真皮成纤维细胞的原代培养物中获得多谱系分化应激耐受细胞（Muse细胞），鉴定其多能性，并检测其分化为黑素细胞的能力。通过免疫组织化学评估人头皮皮肤中SSEA-3阳性细胞的分布，并通过免疫荧光染色确定Oct4、Sox2、Nanog和SSEA-3阳性细胞的分布。分别通过逆转录聚合酶链反应（RT-PCR）分析和蛋白质免疫印迹法测定Muse细胞中Sox2、Oct4、hKlf4和Nanog mRNA及蛋白质的表达水平。这些Muse细胞在分化培养基中分化为黑素细胞。SSEA-3阳性细胞散在于基底膜带和真皮中，在皮脂腺、血管和汗腺以及毛囊外根鞘、真皮乳头和毛球中相对较多。通过用抗SSEA-3抗体进行流式细胞术分选，从头皮真皮成纤维细胞中获得了具有自我更新能力的Muse细胞。RT-PCR、蛋白质免疫印迹和免疫荧光染色结果显示，Muse细胞中Oct4、Nanog、Sox2和Klf4 mRNA及蛋白质的表达水平与其亲代真皮成纤维细胞有显著差异。当在黑素细胞分化培养基中培养时，Muse细胞分化为黑素细胞，且Muse细胞来源的黑素细胞表达黑素细胞特异性标志物HMB45。通过流式细胞术可从头皮真皮成纤维细胞的原代培养物中获得Muse细胞，其具有多能性和自我更新能力，并能在体外分化为黑素细胞。
