Department of Plastic and Reconstructive Surgery, The Ninth People's Hospital affiliated to Shanghai Jiaotong University School of Medicine, 639 Zhi Zao Ju Road, 200011 Shanghai, China.
Skin Res Technol. 2011 Aug;17(3):373-9. doi: 10.1111/j.1600-0846.2011.00510.x. Epub 2011 Feb 21.
Traditional tissue-engineered skin does not produce a satisfactory long-term result because it lacks natural skin pigmentation and leads to discolored cosmetically unpleasing skin that only functions to cover the body of patients. Additionally, the cell sources for tissue-engineered skin are generally derived from normal skin, which is often limited in patients with skin defects.
In this study, hair follicle melanocytes and keratinocytes were isolated from human scalp. The melanocytes were co-cultured with keratinocytes until the second passage and then purified. Purified melanocytes and keratinocytes were seeded onto the chitosan-gelatin membrane for 1 week to construct pigmented tissue-engineered skin. The pigmented skin equivalent was used to resurface the skin defect in nude mice. Four weeks after grafting, skin biopsies were harvested to take hematoxylin and eosin staining and immunohistochemistry staining of Melan-A and HLA-ABC.
Large quantities of purified melanocytes can be obtained with co-culture method. The hematoxylin and eosin staining of repaired skin biopsy demonstrated that the tissue-engineered skin can repair skin defects successfully. Engineered skin contained pigmentation and stained positive for Melan-A and HLA-ABC, which confirmed the presence of melanocytes and its sources were of human origin.
This study demonstrated the possibility of constructing pigmented tissue-engineered skin with human hair follicle-derived keratinocytes and melanocytes, which brings a promising method to make up for the deficiency of traditional tissue-engineered skin and provides an alternative treatment for depigmentation diseases.
传统的组织工程皮肤由于缺乏天然皮肤色素沉着，导致皮肤变色，美容效果不佳，只能起到覆盖患者身体的作用，因此无法取得令人满意的长期效果。此外，组织工程皮肤的细胞来源通常来自正常皮肤，而对于有皮肤缺陷的患者，正常皮肤往往有限。
本研究从人头皮中分离毛囊黑素细胞和成纤维细胞。将黑素细胞与角质形成细胞共培养至第二代，然后进行纯化。纯化后的黑素细胞和成纤维细胞接种到壳聚糖-明胶膜上培养 1 周，构建有色素的组织工程皮肤。将有色素的组织工程皮肤用于裸鼠的皮肤缺损修复。移植后 4 周，采集皮肤活检标本进行苏木精-伊红染色和 Melan-A、HLA-ABC 的免疫组织化学染色。
采用共培养法可获得大量纯化的黑素细胞。修复皮肤活检的苏木精-伊红染色显示，组织工程皮肤可成功修复皮肤缺损。工程皮肤含有色素，Melan-A 和 HLA-ABC 染色阳性，证实了黑素细胞的存在及其来源为人类。
本研究表明，用人毛囊来源的角质形成细胞和成纤维细胞构建有色素的组织工程皮肤是可行的，为弥补传统组织工程皮肤的不足带来了一种有前景的方法，并为脱色素疾病提供了一种替代治疗方法。
