Department of Chemical Engineering, Queen's University, 19 Division Street, Kingston, Ontario, Canada.
Biomaterials. 2012 Jun;33(18):4490-9. doi: 10.1016/j.biomaterials.2012.03.026. Epub 2012 Mar 26.
With the aim of developing a clinically-translatable cell expansion and delivery vehicle for adipose tissue engineering, the adipogenic differentiation of human adipose-derived stem cells (ASCs) was investigated on microcarriers fabricated from human decellularized adipose tissue (DAT). ASCs seeded on the DAT microcarriers and cultured in adipogenic differentiation medium within a low-shear spinner culture system demonstrated high levels of adipogenic differentiation, as measured by the expression of adipogenic genes, glycerol-3-phosphate dehydrogenase (GPDH) enzyme activity, and intracellular lipid accumulation. In contrast, gelatin microcarrier controls did not demonstrate significant adipogenesis, emphasizing the role of the native matrix in mediating ASC differentiation. Interestingly, ASCs cultured on the DAT microcarriers in proliferation medium expressed elevated levels of the adipogenic markers, suggesting that the DAT provided an adipo-inductive substrate for the human ASCs. In vivo testing of the DAT and gelatin microcarriers in a subcutaneous Wistar rat model confirmed injectability and demonstrated stable volume retention over 28 days. Under histological analysis, the DAT microcarriers demonstrated no evidence of immunogenicity or cytotoxicity, with the DAT supporting cellular infiltration and tissue remodeling. Pre-seeding the DAT microcarriers with allogenic rat ASCs enhanced cellularity and angiogenesis within the implant region.
为了开发一种可临床转化的脂肪组织工程细胞扩增和输送载体，研究了人去细胞脂肪组织（DAT）制成的微载体上脂肪源性干细胞（ASCs）的成脂分化。在低剪切搅拌培养系统中，在成脂分化培养基中接种于 DAT 微载体上的 ASC 表现出高水平的成脂分化，这可通过成脂基因的表达、甘油-3-磷酸脱氢酶（GPDH）酶活性和细胞内脂质积累来衡量。相比之下，明胶微载体对照未表现出明显的成脂分化，这强调了天然基质在介导 ASC 分化中的作用。有趣的是，在增殖培养基中培养的 ASC 在 DAT 微载体上表达了高水平的成脂标志物，这表明 DAT 为人类 ASC 提供了一个脂肪诱导的底物。在皮下 Wistar 大鼠模型中对 DAT 和明胶微载体的体内测试证实了可注射性，并在 28 天内表现出稳定的体积保留。在组织学分析中，DAT 微载体未显示出免疫原性或细胞毒性的证据，DAT 支持细胞浸润和组织重塑。用同种异体大鼠 ASC 预先接种 DAT 微载体可增强植入部位的细胞活力和血管生成。
