Dermis-isolated adult stem (DIAS) cells, abundantly available, are attractive for regenerative medicine. Strategies have been devised to isolate and to chondroinduce DIAS cells from various animals. This study aimed to characterize DIAS cells from human abdominal skin (human dermis-isolated adult stem [hDIAS] cells) and to compare and to refine various chondroinduction regimens to form functional neocartilage constructs. The stemness of hDIAS cells was verified (Phase I), three chondroinduction pretreatments were compared (Phase II), and, from these, one regimen was carried forward for refinement in Phase III for improving the mechanical properties of hDIAS cell-derived constructs. Multilineage differentiation and mesenchymal stem cell markers were observed. Among various chondroinduction pretreatments, the nodule formation pretreatment yielded constructs at least 72% larger in diameter, with higher glycosaminoglycan (GAG) content by 44%, compared with other pretreatments. Furthermore, it was found that culturing cells on nontissue culture-treated surfaces yielded constructs (1) on par with constructs derived from aggrecan-coated surfaces and (2) with superior mechanical properties than constructs derived from cells cultured on tissue culture-treated surfaces. After the nodule formation pretreatment, combined supplementation of TGF-β1, IGF-I, and fetal bovine serum significantly enhanced aggregate modulus and shear modulus by 75% and 69%, respectively, over the supplementation by TGF-β1 alone. In summary, human skin-derived DIAS cells are responsive to chondroinduction for forming neocartilage. Furthermore, the mechanical properties of the resultant human constructs can be improved by treatments shown to be efficacious in animal models. Advances made toward tissue-engineering cartilage using animal cells were shown to be applicable to hDIAS cells for cartilage repair and regeneration.

中文翻译：

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用于无支架软骨组织工程的成人真皮干细胞：策略探索

真皮分离的成体干 (DIAS) 细胞数量充足，对再生医学很有吸引力。已经设计了从各种动物中分离和软骨诱导DIAS细胞的策略。本研究旨在表征来自人类腹部皮肤的 DIAS 细胞（人类真皮分离的成人干 [hDIAS] 细胞），并比较和改进各种软骨诱导方案以形成功能性新软骨结构。验证了 hDIAS 细胞的干性（I 阶段），比较了三种软骨诱导预处理（II 阶段），并从中推进了一种方案，以在 III 期进行改进，以改善 hDIAS 细胞衍生结构的机械性能。观察到多向分化和间充质干细胞标志物。在各种软骨诱导预处理中，与其他预处理相比，结节形成预处理产生的构建体直径至少大 72%，糖胺聚糖 (GAG) 含量高 44%。此外，发现在非组织培养处理的表面上培养细胞产生的构建体 (1) 与源自蛋白聚糖涂层表面的构建体相当，(2) 比源自在组织培养处理的表面上培养的细胞的构建体具有更好的机械性能。在结节形成预处理后，与单独补充 TGF-β1 相比，TGF-β1、IGF-I 和胎牛血清的联合补充使聚集模量和剪切模量分别显着提高了 75% 和 69%。总之，人类皮肤来源的 DIAS 细胞对软骨诱导形成新软骨有反应。此外，通过在动物模型中显示有效的治疗，可以改善所得人体构建体的机械性能。使用动物细胞在组织工程软骨方面取得的进展被证明适用于 hDIAS 细胞进行软骨修复和再生。