Oxygen Tension Differentially Influences Osteogenic Differentiation of Human Adipose Stem Cells in 2D and 3D Cultures
Oxygen Tension Differentially Influences Osteogenic Differentiation of Human Adipose Stem Cells in 2D and 3D Cultures
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DOI：
10.1002/jcb.22514
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发表时间：
2010-05-01
影响因子：
4
通讯作者：
Leach, J. Kent
中科院分区：
文献类型：
--
作者：
He, Jiawei;Genetos, Damian C.;Leach, J. Kent
骨缺损通常遭受由与损伤或疾病相关的受损血管形成引起的缺氧微环境。脂肪干细胞（ASC）代表了一个有前途的细胞群，通过向成骨谱系分化或分泌营养因子来刺激骨骼修复。然而，ASCs对低氧微环境的成骨或营养反应知之甚少。此外，缺乏ASCs对缺氧的2D和3D反应之间的直接比较。因此，我们在培养4周的2D和3D中表征了低氧（1%）、常氧（5%）和大气（21%）氧张力下人ASC的成骨和血管生成潜力。我们检测到最大的碱性磷酸酶活性和细胞外钙沉积在2D和3D下21%的氧气培养的细胞，酶活性的降低对应于氧张力的降低。在1%氧气中培养的ASCs在4周内比在其他条件下培养的细胞分泌更多的血管内皮生长因子（VEGF），在2D中培养的细胞比在3D中培养的细胞以更持续的方式分泌VEGF。成骨标志物的表达在不同的氧条件下显示时间变化，在3D中表达峰值出现较早。此外，在1%和5%氧气条件下，2D培养物中大多数成骨标志物的增加显著高于3D培养物。这些结果表明，氧气，结合维度，影响ASCs的分化程序的时间。这些发现为ASCs在骨修复中的应用提供了新的见解，同时强调了培养微环境的重要性。110：87-96，2010. (C)2010年威利-利斯。Inc.
Skeletal defects commonly suffer from poor oxygen microenvironments resulting from compromised vascularization associated with injury or disease. Adipose stem cells (ASCs) represent a promising cell population for stimulating skeletal repair by differentiating toward the osteogenic lineage or by secreting trophic factors. However, the osteogenic or trophic response of ASCs to reduced oxygen microenvironments is poorly understood. Moreover, a direct comparison between 2D and 3D response of ASCs to hypoxia is lacking. Thus, we characterized the osteogenic and angiogenic potential of human ASCs under hypoxic (1%), normoxic (5%), and atmospheric (21%) oxygen tensions in both 2D and 3D over 4 weeks in culture. We detected greatest alkaline phosphatase activity and extracellular calcium deposition in cells cultured in both 2D and 3D under 21% oxygen, and reductions in enzyme activity corresponded to reductions in oxygen tension. ASCs cultured in 1% oxygen secreted more vascular endothelial growth factor (VEGF) over the 4-week period than cells cultured in other conditions, with cells cultured in 2D secreting VEGF in a more sustained manner than those in 3D. Expression of osteogenic markers revealed temporal changes under different oxygen conditions with peak expression occurring earlier in 3D. In addition, the increase of most osteogenic markers was significantly higher in 2D compared to 3D cultures at 1% and 5% oxygen. These results suggest that oxygen, in conjunction with dimensionality, affects the timing of the differentiation program in ASCs. These findings offer new insights for the use of ASCs in bone repair while emphasizing the importance of the culture microenvironment J. Cell. Biochem. 110: 87-96, 2010. (C) 2010 Wiley-Liss. Inc.