Mixture of Fibroblasts and Adipose Tissue-Derived Stem Cells Can Improve Epidermal Morphogenesis of Tissue-Engineered Skin
Mixture of Fibroblasts and Adipose Tissue-Derived Stem Cells Can Improve Epidermal Morphogenesis of Tissue-Engineered Skin
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DOI：
10.1159/000324921
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发表时间：
2011-04
影响因子：
2.7
通讯作者：
W. Lu;Jinhua Yu;Yong-jie Zhang;K. Ji;Yalei Zhou;Yuan Li;Z. Deng;Yan Jin
中科院分区：
文献类型：
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作者：
W. Lu;Jinhua Yu;Yong-jie Zhang;K. Ji;Yalei Zhou;Yuan Li;Z. Deng;Yan Jin
许多研究表明，相邻间充质细胞的类型可以影响双层组织工程皮肤的表皮形态发生。然而，不同间充质细胞类型的混合物是否可以改善生物工程皮肤的表皮形态发生仍然未知。在这项研究中，角质形成细胞，真皮成纤维细胞和脂肪组织来源的干细胞（ADSCs）的分离和纯化，从人类皮肤和皮下脂肪。MTT法显示，真皮成纤维细胞与脂肪干细胞按1：1比例混合培养的条件培养液对角质形成细胞增殖的促进作用上级单独培养的成纤维细胞或脂肪干细胞。此外，ELISA结果显示，混合成纤维细胞/ADSC组中的人肝细胞生长因子和角质形成细胞生长因子（也称为FGF 7）的细胞因子水平高于ADSC或真皮成纤维细胞组中的那些。为了研究混合成纤维细胞和脂肪干细胞对表皮形态发生的潜在作用，应用三维组织工程皮肤系统。组织学分析表明，角质形成细胞在成纤维细胞和脂肪干细胞的混合物中广泛增殖，并形成具有良好分化结构的厚表皮层。成纤维细胞和脂肪干细胞混合组的双层组织工程皮肤基底层上表达表皮分化标志物角蛋白10。透射电镜观察到新生表皮中存在桥粒和半桥粒。总之，这些发现首次揭示了双层组织工程皮肤中成纤维细胞和ADSC的混合物可以改善表皮形态发生。
Many studies demonstrate that the type of adjacent mesenchymal cells can affect epidermal morphogenesis of bilayered tissue-engineered skin. However, whether a mixture of different mesenchymal cell types can improve epidermal morphogenesis of bioengineered skin remains unknown. In this study, keratinocytes, dermal fibroblasts and adipose tissue-derived stem cells (ADSCs) were isolated and purified from human skin and subcutaneous fat. Conditioned medium generated from a mixture of dermal fibroblasts and ADSCs at the ratio of 1:1 was superior to that from fibroblasts or ADSCs alone in promoting keratinocyte proliferation, as indicated by MTT assay. Furthermore, ELISA results showed that the cytokine levels of human hepatocyte growth factor and keratinocyte growth factor (also known as FGF7) in the mixed fibroblasts/ADSC group were higher than those in the ADSC or dermal fibroblasts group. To examine the potential roles of mixed fibroblasts and ADSCs on epidermal morphogenesis, a three-dimensional tissue engineered skin system was applied. Histological analyses demonstrated that keratinocytes proliferated extensively over the mixture of fibroblasts and ADSCs, and formed a thick epidermal layer with well-differentiated structures. Keratin 10 (epidermal differentiation marker) was expressed in the suprabasal layer of bilayered tissue-engineered skin in the mixed fibroblasts and ADSCs group. Desmosomes and hemidesmosomes were detected in the newly formed epidermis by transmission electron microscopy analysis. Together, these findings revealed for the first time that a mixture of fibroblasts and ADSCs in bilayered tissue-engineered skin can improve epidermal morphogenesis.