College of Pharmacy and Research Institute of Pharmaceutical Sciences, Seoul National University, Seoul 151-742, South Korea.
J Biosci Bioeng. 2011 Oct;112(4):402-8. doi: 10.1016/j.jbiosc.2011.06.018. Epub 2011 Jul 30.
Human adipose-derived mesenchymal stem cells (AD-MSCs) attracted much interest as a promising alternative to autologous chondrocytes and bone marrow-derived mesenchymal stem cells for cartilage regeneration. Developing a suitable culture technique to direct AD-MSCs into the chondrogenic lineage could be a crucial prerequisite for the cartilage defect repair application of AD-MSCs. Herein, we prepared the PEGDG-crosslinked porous three-dimensional (3D) hyaluronic acid (HA) scaffold and evaluated for its feasibility to induce proliferation and chondrogenic differentiation of the AD-MSCs. In addition, the effect of bone-morphogenetic protein-2 (BMP-2) and platelet-derived growth factor (PDGF) on chondrogenic differentiation was further investigated. Proliferation and chondrogenic differentiation were evaluated by cell morphology, DNA contents, s-GAG contents, and level of mRNA expression of relevant marker genes. When cultured with reference chondrogenic medium (RCM; serum-free DMEM-HG supplemented with 10 ng/mL of transforming growth factor-β1 (TGF-β1), 50 nM ascorbate, 100 nM dexamethasone, and 5 μg/mL of ITS), better proliferation and chondrogenic differentiation of AD-MSCs were obtained in the 3D HA scaffold culture as compared to the micromass culture, a standard 3D culture system. Moreover, the level of chondrogenic differentiation of AD-MSCs in the HA scaffold-RCM culture system was further increased by BMP-2, and decreased by PDGF. These results suggested that the HA scaffold with RCM was a promising chondrogenic culture system of AD-MSCs, and that BMP-2 could potentially serve as a chondrogenic supplement for AD-MSCs. However, PDGF was determined to be an inappropriate supplement based on its inhibition of the chondrogenic differentiation of AD-MSCs.
人脂肪间充质干细胞（AD-MSCs）作为一种有前途的替代物，吸引了人们的极大兴趣，可用于软骨再生，替代自体软骨细胞和骨髓间充质干细胞。开发合适的培养技术，将 AD-MSCs 定向诱导为软骨细胞谱系，可能是 AD-MSCs 软骨缺损修复应用的关键前提。本文中，我们制备了 PEGDG 交联多孔三维（3D）透明质酸（HA）支架，并评估了其诱导 AD-MSCs 增殖和软骨分化的可行性。此外，还进一步研究了骨形态发生蛋白-2（BMP-2）和血小板衍生生长因子（PDGF）对软骨分化的影响。通过细胞形态、DNA 含量、s-GAG 含量和相关标记基因的 mRNA 表达水平评估增殖和软骨分化。当与参考软骨形成培养基（RCM；无血清 DMEM-HG 补充有 10ng/ml 转化生长因子-β1（TGF-β1）、50nM 抗坏血酸、100nM 地塞米松和 5μg/ml ITS）一起培养时，与微团培养相比，AD-MSCs 在 3D HA 支架培养中获得了更好的增殖和软骨分化，微团培养是一种标准的 3D 培养系统。此外，在 HA 支架-RCM 培养系统中，BMP-2 进一步增加了 AD-MSCs 的软骨分化水平，而 PDGF 则降低了 AD-MSCs 的软骨分化水平。这些结果表明，含有 RCM 的 HA 支架是一种有前途的 AD-MSCs 软骨形成培养系统，BMP-2 可能作为 AD-MSCs 的软骨形成补充物。然而，根据 PDGF 抑制 AD-MSCs 软骨分化的作用，确定 PDGF 是不合适的补充物。
