Upregulation of Adipogenesis and Chondrogenesis in MSC Serum-Free Culture
Upregulation of Adipogenesis and Chondrogenesis in MSC Serum-Free Culture
复制标题
DOI：
10.3727/215517911x575984
复制
发表时间：
2011-04-01
期刊：
影响因子：
--
通讯作者：
Lee, Eng Hin
中科院分区：
文献类型：
--
作者：
Ho, Saey Tuan Barnabas;Tanavde, Vivek Madhukar;Lee, Eng Hin
无血清培养基已被证明可有效扩增间充质干细胞 (MSC)。然而，其影响可能超出细胞扩增范围，因为细胞的分化潜力可能会被改变，从而影响其下游应用的功效。人们对后者知之甚少，这促使人们评估无血清制剂对软骨形成、脂肪形成的影响。和 MSC 的成骨潜力。该培养基由 Knockout (TM) 血清替代品 (KSR) 和生长因子混合物以及胶原蛋白或纤连蛋白涂层组成。选择胶原蛋白涂层是因为它促进一致的细胞附着。与胎牛血清 (FRS) 对照相比，无血清培养基中的细胞增殖在第 1 代时增强。在第 3 代时，两组中观察到相似水平的表面标志物，而无血清培养物中的 CD90 和 CD73 略有减少。在分化条件下筛选培养物，发现无血清培养基中的软骨形成潜力得到更好的维持，糖胺聚糖 (GAG) 和胶原蛋白 II 的表达更高。 FBS 组的软骨形成存在缺陷，这是由于动物血清固有的不一致所致。无血清组的脂肪生成增强，PPARG 表达和脂质积累更高。 FBS 组和无血清组的成骨矿化水平相似，但后者的 I 型胶原蛋白基因表达受到抑制。这最初是在扩张期间观察到的。这些观察结果归因于无血清制剂中存在的细胞因子触发的信号级联反应以及与胶原蛋白底物的相互作用。无血清培养基有助于分别维持和增强 MSC 的软骨形成和脂肪形成潜力。这一优势可用于软骨和脂肪组织工程的治疗应用。
Serum-free media have been shown to be effective in the expansion of mesenchymal stem cells (MSCs). However, the effects may go beyond cell expansion as the differentiation potentials of the cells may be modified, thus influencing their efficacy for downstream applications. The latter is poorly understood, and this has prompted an evaluation of the influence of a serum-free formulation on the chondrogenic, adipogenic. and osteogenic potential of MSCs. The media consisted of Knockout (TM) Serum Replacement (KSR) with a cocktail of growth factors coupled with either collagen or fibronectin coatings. Collagen coating was selected as it promoted consistent cellular attachment. When compared against fetal bovine serum (FRS) controls, cell proliferation in the serum-free media was enhanced at passage 1. Similar levels of surface markers were observed in the two groups with a slight reduction in CD90 and CD73 in the serum-free culture at passage 3. The cultures were screened under differentiation conditions and a better maintenance of the chondrogenic potential was noted in the serum-free media with higher expressions of glycoaminoglycans (GAGs) and collagen II. Chondrogenesis was deficient in the FBS group and this was attributed to the inherent inconsistency of animal serum. Adipogenesis was enhanced in the serum-free group with a higher PPARG expression and lipid accumulation. Similar levels of osteogenic mineralization was noted in the FBS and serum-free groups but collagen I gene expression was suppressed in the latter. This was initially observed during expansion. These observations were attributed to the signaling cascades triggered by the cytokines presented in the serum-free formulation and the interaction with the collagen substrate. The serum-free media helps to maintain and enhance the chondrogenic and adipogenic potentials of the MSCs, respectively. This advantage can be exploited for therapeutic applications in cartilage and adipose tissue engineering.