Decorin moieties tethered into PEG networks induce chondrogenesis of human mesenchymal stem cells.
Decorin moieties tethered into PEG networks induce chondrogenesis of human mesenchymal stem cells.
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DOI：
10.1002/jbm.a.32112
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发表时间：
2009-08
期刊：
影响因子：
--
通讯作者：
Anseth KS
中科院分区：
文献类型：
--
作者：
Salinas CN;Anseth KS
一种通用的方法来制造聚乙二醇-肽共聚物凝胶被用来设计壁龛，以促进人骨髓间充质干细胞（hMSCs）的软骨分化。选择序列RGD和KLER作为基序，通过硫醇-丙烯酸酯聚合来修饰PEG凝胶。已知KLER序列（来自核心蛋白聚糖蛋白的结合位点）与II型胶原蛋白强烈结合并且负责基质组织，而RGD促进包囊细胞的一般存活。在存在或不存在5 mM CKLERG的情况下，将hMSC以2 × 106个细胞/mL包封到含有1 mM CRGDSG的10 wt % PEG凝胶中。一个加扰序列作为对照。将凝胶在含有5 ng/mL TGFβ1的对照和软骨形成培养基中培养6周。在不同时间点分析细胞/凝胶构建体的糖胺聚糖（GAG）含量、II型胶原沉积、免疫染色和基因分析。在软骨形成培养物中14天后，RGDS和KLER功能化凝胶中的细胞产生的GAG/细胞是仅含有RGD的凝胶中的细胞的2.5倍。到第28天，软骨形成KLER凝胶内的hMSC产生的羟脯氨酸比第0天高27倍，而单独使用RGDS的软骨形成培养物中的细胞产生的羟脯氨酸是初始的两倍。免疫染色图像表明，Col II在KLER衍生的凝胶中比其他凝胶更占优势，并且通过II型胶原和聚集蛋白聚糖表达的RT-PCR分析进一步支持在含有KLER的凝胶中增强的软骨形成分化。总的来说，这些结果证明了基质结合肽的掺入如何与诱导软骨形成分化和软骨特异性ECM沉积的hMSC相互作用。
A versatile approach to fabricate PEG-peptide copolymer gels was utilized to design niches to promote chondrogenic differentiation of human mesenchymal stem cells (hMSCs). The sequences RGD and KLER were chosen as motifs to modify PEG gels through a thiol-acrylate polymerization. The KLER sequence, a binding site from decorin protein, is known to bind strongly to collagen type II and is responsible for matrix organization, while RGD promotes general survival of encapsulated cells. hMSCs were encapsulated at 2 × 106 cells/mL into 10 wt % PEG gels with 1 mM CRGDSG in the presence or absence of 5 mM CKLERG. A scrambled sequence served as a control. The gels were cultured in control and chondrogenic media, containing 5 ng/mL TGFβ1 over a 6-week period. Cell/gel constructs were analyzed at various time points for glycosaminoglycan (GAG) content, type II collagen deposition, immunostaining, and gene analysis. After 14 days in chondrogenic cultures, cells in RGDS and KLER functionalized gels produced 2.5 times as much GAG/cell as those in gels containing only RGD. By day 28, hMSCs within the chondrogenic KLER gels produced 27-fold higher hydroxyproline than that of day 0, whereas cells in chondrogenic culture with RGDS alone produced twofold of initial. Immunostained images indicated that col II was more predominant in the KLER-derivatized gels than others, and enhanced chondrogenic differentiation in KLER containing gels was further supported by RT-PCR analysis of type II collagen and aggrecan expression. Collectively, these results demonstrate how incorporation of matrix-binding peptide interacts with hMSCs inducing chondrogenic differentiation and cartilage-specific ECM deposition.
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影响因子：
14
作者：
Mann, BK;Gobin, AS;West, JL
通讯作者：
West, JL
DOI：
10.1016/0304-4165(86)90306-5
发表时间：
1986-09-04
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
FARNDALE, RW;BUTTLE, DJ;BARRETT, AJ
通讯作者：
BARRETT, AJ
影响因子：
14
作者：
Mann, BK;Schmedlen, RH;West, JL
通讯作者：
West, JL
DOI：
10.1073/pnas.83.20.7683
发表时间：
1986-10-01
影响因子：
11.1
作者：
KRUSIUS, T;RUOSLAHTI, E
通讯作者：
RUOSLAHTI, E
影响因子：
14
作者：
Lee, CR;Grodzinsky, AJ;Spector, M
通讯作者：
Spector, M