Here, the in vitro engineering of a cartilage-like tissue by using decellularized extracellular matrix scaffold (hECM) seeded with human adipose stem cells (hASCs) which can both be isolated from the human waste adipose tissue is described. Cell-free, highly fibrous and porous hECM was produced using a protocol containing physical (homogenization, centrifugation, molding) and chemical (crosslinking) treatments, characterized by SEM, histochemistry, immunohistochemistry and in vitro cell interaction study. A construct of hECM seeded with hASCs was cultured in chondrogenic medium (with TGF-β3 and BMP-6) for 42 days. SEM and histology showed that the biological scaffold was highly porous and had a compact structure suitable for handling and subsequent cell culture stages. Cells successfully integrated into the scaffold and had good cellular viability and continuity to proliferate. Constructs showed the formation of cartilage-like tissue with the synthesis of cartilage-specific proteins, Collagen type II and Aggrecan. Dimethylmethylene blue dye binding assay demonstrated that the GAG content of the constructs was in tendency to increase with time confirming chondrogenic differentiation of hASCs. The results support that human waste adipose tissue is an important source for decellularized hECM as well as stem cells, and adipose hECM scaffold provides a suitable environment for chondrogenic differentiation of hASCs.

中文翻译：

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用于软骨组织工程的来自自体人脂肪组织的脱细胞生物支架和干细胞

在这里，描述了通过使用接种人类脂肪干细胞 (hASCs) 的脱细胞细胞外基质支架 (hECM) 进行软骨样组织的体外工程，这些细胞都可以从人类废物脂肪组织中分离出来。无细胞、高纤维和多孔 hECM 是使用包含物理（均质化、离心、成型）和化学（交联）处理的协议生产的，其特征在于 SEM、组织化学、免疫组织化学和体外细胞相互作用研究。用 hASCs 接种的 hECM 构建体在软骨形成培养基（含有 TGF-β3 和 BMP-6）中培养 42 天。SEM 和组织学表明，生物支架是高度多孔的，并且具有适合处理和后续细胞培养阶段的紧凑结构。细胞成功整合到支架中，并具有良好的细胞活力和增殖连续性。构建体显示软骨样组织的形成与软骨特异性蛋白质、II 型胶原蛋白和聚集蛋白聚糖的合成。二甲基亚甲基蓝染料结合测定表明，构建体的 GAG 含量有随时间增加的趋势，证实了 hASC 的软骨分化。结果支持人类废弃脂肪组织是脱细胞 hECM 和干细胞的重要来源，脂肪 hECM 支架为 hASC 的软骨分化提供了合适的环境。二甲基亚甲基蓝染料结合测定表明，构建体的 GAG 含量有随时间增加的趋势，证实了 hASC 的软骨分化。结果支持人类废弃脂肪组织是脱细胞 hECM 和干细胞的重要来源，脂肪 hECM 支架为 hASC 的软骨分化提供了合适的环境。二甲基亚甲基蓝染料结合测定表明，构建体的 GAG 含量有随时间增加的趋势，证实了 hASC 的软骨分化。结果支持人类废弃脂肪组织是脱细胞 hECM 和干细胞的重要来源，脂肪 hECM 支架为 hASC 的软骨分化提供了合适的环境。