Effects of resveratrol on enrichment of adipose-derived stem cells and their differentiation to osteoblasts in two-and three-dimensional cultures
Effects of resveratrol on enrichment of adipose-derived stem cells and their differentiation to osteoblasts in two-and three-dimensional cultures
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DOI：
10.1002/term.513
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发表时间：
2012-12-01
影响因子：
3.3
通讯作者：
Boyan, Barbara D.
中科院分区：
文献类型：
--
作者：
Erdman, Christopher P.;Dosier, Christopher R.;Boyan, Barbara D.
本研究的目的是开发一种从皮下脂肪中分离多能脂肪间充质干细胞（ASC）和骨祖细胞（OPC）的方法。从大鼠腹股沟脂肪中去除成熟脂肪细胞和造血细胞后，通过其对塑料的附着、表面标志物特征（CD 271+、CD 73+和CD 45）以及分化成脂肪细胞、软骨细胞和成骨细胞的能力来验证剩余细胞群中的ASC。OPCs定义为E11+和OCN+。贴壁细胞培养在生长培养基（GM）或成骨培养基（OM）和白藜芦醇（0，12.5，和25 μ m）处理7？通过流式细胞术评估ASC和OPC。成骨潜能在二维（2D）培养中作为碱性磷酸酶特异性活性和骨钙素产生的函数来确定。此外，细胞接种到三维（3D）聚-e-己内酯支架和动态条件下培养，矿化定量微CT在4，8和12？周白藜芦醇可增加GM和OM中ASCs的数量，但仅增加GM中OPCs的数量。在这两种媒体类型白藜芦醇增加碱性磷酸酶活性和骨钙素水平。在3D培养中，白藜芦醇处理的细胞在早期时间点显著增加矿化基质体积。白藜芦醇通过富集ASC和OPC群体并增强成骨分化对贴壁细胞产生双相作用。白藜芦醇预处理诱导更多的矿化在较早的时间点，并代表了临床上可行的技术，骨科和牙科应用自体干细胞治疗。版权所有（C）2012约翰威利父子有限公司
The goal of this study was to develop a method for increasing the yield of multipotent adipose-derived mesenchymal stem cells (ASCs) and osteoprogenitor cells (OPCs) from subcutaneous fat. After removing mature adipocytes and haematopoietic cells from rat inguinal fat, ASCs in the remaining cell population were verified by their attachment to plastic, surface marker profile (CD271+, CD73+ and CD45) and ability to differentiate into adipocytes, chondrocytes and osteoblasts. OPCs were defined as E11+ and OCN+. Adherent cells were cultured in growth medium (GM) or osteogenic medium (OM) and treated with resveratrol (0, 12.5, and 25 mu m) for 7?days; ASCs and OPCs were assessed by flow cytometry. Osteogenic potential was determined in two-dimensional (2D) cultures as a function of alkaline phosphatase-specific activity and osteocalcin production. In addition, cells were seeded onto three-dimensional (3D) poly-e-caprolactone scaffolds and cultured under dynamic conditions; mineralization was quantified by micro-CT at 4, 8 and 12?weeks. Resveratrol increased the percentage of ASCs in the population (population%) and number of ASCs in both GM and OM, but increased only the number of OPCs in GM. In both media types resveratrol increased alkaline phosphatase activity and osteocalcin levels. In 3D cultures, resveratrol-treated cells significantly increased mineralized matrix volume at early time points. Resveratrol exerted a biphasic effect on adherent cells by enriching the ASC and OPC populations and enhancing osteogenic differentiation. Resveratrol pretreatment induced more mineralization at earlier time points and represents a clinically viable technique for orthopaedic and dental applications for autologous stem cell therapy. Copyright (C) 2012 John Wiley & Sons, Ltd.