The guidance of human mesenchymal stem cell differentiation in vitro by controlled modifications to the cell substrate
The guidance of human mesenchymal stem cell differentiation in vitro by controlled modifications to the cell substrate
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DOI：
10.1016/j.biomaterials.2006.05.001
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发表时间：
2006-09-01
期刊：
影响因子：
14
通讯作者：
Hunt, John A.
中科院分区：
文献类型：
--
作者：
Curran, Judith M.;Chen, Rui;Hunt, John A.
骨髓间充质干细胞（MSC）的行为和分化的材料驱动控制是一个非常令人兴奋的可能性。本研究的目的是使用硅烷改性的表面来控制MSC的粘附和分化在体外和评估使用这些技术来控制MSC的行为在基础和刺激条件。产生一系列表征的清洁玻璃硅烷改性表面，甲基（-CH 3）、氨基（-NH 2）、硅烷（-SH）、羟基（-OH）和羧基（-COOH），并与人MSC接触培养，结合清洁玻璃（TAAB）对照，在基础、软骨形成和成骨刺激培养基中培养长达28天。使用荧光免疫组织化学（胶原蛋白I、II、骨钙蛋白、CBFA 1）和实时聚合酶链反应（PCR）（胶原蛋白I、II、骨钙蛋白、骨桥蛋白、骨连接蛋白、CBFA 1和Sox 9）分析样本的活细胞粘附水平、形态和各种分化和转录标志物的产生。对结果的分析表明，材料的范围可分为三个不同的类别。首先，-TAAB控制和-CH 3表面保持MSC表型;其次，-NH 2和-SH-修饰的表面在生物刺激的存在和不存在下促进和维持骨生成。这些表面在任何测试条件下都不支持长期软骨形成。最后，-OH和-COOH-改性的表面促进和维持软骨形成的基础和软骨刺激条件下，但不支持成骨。这些结果表明，复杂的材料特性，如表面化学和能量，可以影响MSC的体外行为。这些结果不仅在促进组织工程构建的效率方面有意义，而且在MSC分离、维持和扩增的更广泛领域也有意义。(c)2006爱思唯尔有限公司保留所有权利。
Material-driven control of bone-marrow-derived mesenchymal stem cell (MSC) behaviour and differentiation is a very exciting possibility. The aim of this study was to use silane-modified surfaces to control MSC adhesion and differentiation in vitro and evaluate the use of such techniques to control MSC behaviour both in basal and stimulated conditions. A range of characterised clean glass silane-modified surfaces, methyl (-CH3), amino (-NH2), silane (-SH), hydroxyl (-OH) and carboxyl (-COOH), were produced and cultured in contact with human MSC, in conjunction with a clean glass (TAAB) control, for time periods up to 28 days in basal, chondrogenic and osteogenic stimulated media. The samples were analysed for levels of viable cell adhesion, morphology and the production of various differentiation and transcription markers using both fluorescent immunohistochemistry (collagen I, II, osteocalcin, CBFA1) and real-time polymerase chain reaction (PCR) (collagen I, II, osteocalcin, osteopontin, osteonectin, CBFA1 and Sox 9). Analysis of the results demonstrated that the range of materials could be broken down into three distinct categories. Firstly, the -TAAB control and -CH3 surfaces maintained the MSC phenotype; secondly, the -NH2 and -SH-modified surfaces promoted and maintained osteogenesis both in the presence and absence of biological stimuli. These surfaces did not support long-term chondrogenesis under any test conditions. Finally, the -OH and -COOH-modified surfaces promoted and maintained chondrogenesis under both basal and chondrogenic stimulated conditions, but did not support osteogenesis. These results demonstrate that intricate material properties such as surface chemistry and energy can influence MSC behaviour in vitro. These results have implications not only in promoting the efficiency of tissue-engineered constructs, but also to the wider field of MSC isolation, maintenance and expansion. (c) 2006 Elsevier Ltd. All rights reserved.