琼脂糖培养中人骨髓间充质干细胞的软骨形成。

C-Y Charles Huang, Paul M Reuben, Gianluca D'Ippolito, Paul C Schiller, Herman S Cheung
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引用次数: 163

摘要

来源于人骨髓的间充质干细胞(hBM-MSCs)可以分化为软骨细胞,用于潜在的关节软骨损伤治疗。为了评估琼脂糖凝胶作为hBM-MSCs软骨形成的支持材料,本研究在琼脂糖培养中检测了hBM-MSCs的软骨形成。采用颗粒培养来确认琼脂糖培养中使用的hBM-MSCs的成软骨潜能。hBM-MSCs在2%琼脂糖结构中播种,初始细胞播种密度为3、6和9 × 10(6)个细胞/ml,而每个颗粒使用2.5 × 10(5)个细胞形成。在含有转化生长因子β 3 (tgf - β 3)的培养基中,通过培养细胞琼脂糖构建物和微球21天诱导hBM-MSCs的软骨形成。逆转录-聚合酶链反应分析显示,琼脂糖和颗粒培养的hBM-MSCs在tgf - β 3存在下表达了II型胶原和聚集蛋白的软骨形成标志物。通过组织学和免疫组织化学评估,琼脂糖和颗粒培养中均检测到软骨特异性大分子的沉积。琼脂糖凝胶中hBM-MSCs的软骨形成与初始细胞播种密度直接相关,初始细胞播种密度高的细胞琼脂糖构建体表现出更多的软骨特异性基因表达。本研究为今后琼脂糖培养hBM-MSCs的软骨形成研究奠定了基础模型。
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Chondrogenesis of human bone marrow-derived mesenchymal stem cells in agarose culture.

Mesenchymal stem cells derived from human bone marrow (hBM-MSCs) can differentiate into chondrogenic cells for the potential treatment of injured articular cartilage. To evaluate agarose gels as a supportive material for chondrogenesis of hBM-MSCs, this study examined chondrogenesis of hBM-MSCs in the agarose cultures. Pellet cultures were employed to confirm the chondrogenic potential of the hBM-MSCs that were used in agarose cultures. The hBM-MSCs were seeded in 2% agarose constructs at the initial cell-seeding densities of 3, 6, and 9 x 10(6) cells/ml while each of pellets was formed using 2.5 x 10(5) cells. Chondrogenesis of hBM-MSCs was induced by culturing cell-agarose constructs and pellets for 21 days in the presence of a defined medium containing transforming growth factor beta3 (TGF-beta3). The analysis of reverse transcription-polymerase chain reaction showed that hBM-MSCs of agarose and pellet cultures expressed the chondrogenic markers of collagen type II and aggrecan in the presence of TGF-beta3. The deposition of cartilage-specific macromolecules was detected in both agarose and pellet cultures by histological and immunohistochemical assessments. Chondrogenesis of hBM-MSCs in agarose gels directly correlated with the initial cell-seeding density, with the cell-agarose constructs of higher initial cell-seeding density exhibiting more cartilage-specific gene expressions. This study establishes a basic model for future studies on chondrogenesis of hBM-MSCs using the agarose cultures.

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