Growth Differentiation Factor-5 Enhances In Vitro Mesenchymal Stromal Cell Chondrogenesis and Hypertrophy
Growth Differentiation Factor-5 Enhances In Vitro Mesenchymal Stromal Cell Chondrogenesis and Hypertrophy
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DOI：
10.1089/scd.2012.0282
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发表时间：
2013-07-01
影响因子：
4
通讯作者：
Barry, Frank
中科院分区：
文献类型：
--
作者：
Coleman, Cynthia M.;Vaughan, Erin E.;Barry, Frank
成人骨髓间充质干细胞（MSCs）的再生潜力已被广泛研究，在关节炎疾病和局灶性软骨缺损的设置。MSC的体外成软骨分化通常通过广泛使用的团块培养系统来完成，其中MSC保持在高密度团块中以模拟发育期间的间充质凝聚。补充软骨形成MSC颗粒培养物与生长分化因子-5（GDF-5），一个高度调节的基因在软骨内骨化（EO）的软骨形成阶段，在这里进行了研究的假设下，GDF-5将增强骨髓间充质干细胞的软骨分化，从而支持他们进入骨化。用重组人GDF-5蛋白补充成软骨MSC颗粒显著增强MSC成软骨分化，如通过增强的II型胶原和硫酸化糖胺聚糖（GAG）掺入细胞外基质所证明的。当与单独用TGF-β 3处理的颗粒相比时，在用GDF-5和转化生长因子（TGF）-β 3处理的颗粒中观察到P-SMAD 1-5-8增加，这通过软骨形成颗粒提取物的免疫染色和蛋白质印迹分析来证明。碱性磷酸酶，I型和X型胶原蛋白，骨桥蛋白分泌的同时增加，表明这些文化的过度肥大。总之，这些数据支持GDF-5作为EO前体用于增强MSC软骨形成分化和肥大。
The regenerative potential for adult bone marrow-derived mesenchymal stromal cells (MSCs) has been extensively investigated in the setting of arthritic disease and focal cartilage defects. In vitro chondrogenic differentiation of MSCs is regularly accomplished by the widely used pellet culture system where MSCs are maintained in high-density pellets to mimic mesenchymal condensation during development. Supplementation of chondrogenic MSC pellet cultures with growth differentiation factor-5 (GDF-5), a highly regulated gene in the chondrogenic phase of endochondral ossification (EO), was investigated here under the hypothesis that GDF-5 will enhance the chondrogenic differentiation of MSCs, thereby supporting their entry into ossification. The supplementation of chondrogenic MSC pellets with the recombinant human GDF-5 protein significantly enhanced MSC chondrogenic differentiation, as demonstrated by enhanced collagen type II and sulfated glycosaminoglycan (GAG) incorporation into the extracellular matrix. Increased P-SMADs 1-5-8 were observed in pellets treated with GDF-5 and transforming growth factor (TGF)-beta 3 when compared to the pellets treated with TGF-beta 3 alone, demonstrated by immunostaining and western blot analysis of the chondrogenic pellet extract. A concurrent increase in alkaline phosphatase, collagen types I and X, and osteopontin secretion indicated a transition of these cultures to hypertrophy. Together, these data support the application of GDF-5 to enhance MSC chondrogenic differentiation and hypertrophy as a precursor to EO.