Tissue-specific bioactivity of soluble tendon-derived and cartilage-derived extracellular matrices on adult mesenchymal stem cells.
Tissue-specific bioactivity of soluble tendon-derived and cartilage-derived extracellular matrices on adult mesenchymal stem cells.
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DOI：
10.1186/s13287-017-0580-8
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发表时间：
2017-06-05
影响因子：
7.5
通讯作者：
Tuan RS
中科院分区：
文献类型：
--
作者：
Rothrauff BB;Yang G;Tuan RS
由组织来源的细胞外基质（ECM）组成的生物支架可以促进同源（即，组织特异性）细胞分化。最近，来自脱细胞肌腱和软骨的可溶性ECM被推广为组织特异性生物材料，但溶解后是否保留组织特异性生物活性尚不清楚。本研究探讨了可溶性脱细胞肌腱和软骨ECM对人骨髓间充质干细胞（MSC）的组织特异性生物活性，这些细胞在不同的培养微环境中存在，包括二维（2D）组织培养塑料，对齐的静电纺丝纳米纤维，细胞团和细胞接种的光交联水凝胶。使用已建立的方法将肌腱和软骨ECM脱细胞，并通过胃蛋白酶消化或尿素提取溶解。通过在2D组织培养塑料上培养的人MSC的基础培养基中补充可溶性ECM对细胞增殖和分化的影响进行了初步探索。在随后的实验中，将MSC培养在对齐的电纺纳米纤维、ascell小球上，或封装在可光交联的甲基丙烯酸酯化明胶（GelMA）水凝胶内。添加尿素提取的肌腱和软骨ECM作为补充剂。胃蛋白酶消化的ECM不促进人MSC的同源分化，无论是作为培养基补充物还是三维（3D）水凝胶提供。相比之下，尿素提取的ECM倾向于促进在2D和3D微环境中培养的MSC的组织特异性分化。小分子TGF-β信号传导抑制剂SB-431542的应用在很大程度上否定了由肌腱和软骨ECM介导的组织特异性基因表达模式。这表明内源性TGF-β的作用是必需的，但不足以赋予尿素提取的ECM的组织特异性生物活性。当将尿素提取的软骨ECM掺入光可固化GelMA水凝胶中时，其独立地增强了包封的MSC中的软骨形成，并且在培养基中补充TGF-β后显示出附加的原软骨形成。脱细胞肌腱和软骨的尿素提取ECM组分是能够增强成体干细胞的组织特异性分化的可溶性补充剂。本文的在线版本（doi：10.1186/s13287-017-0580-8）包含补充材料，可供授权用户使用。
Biological scaffolds composed of tissue-derived extracellular matrix (ECM) can promote homologous (i.e., tissue-specific) cell differentiation through preservation of biophysical and biochemical motifs found in native tissues. Solubilized ECMs derived from decellularized tendon and cartilage have recently been promoted as tissue-specific biomaterials, but whether tissue-specific bioactivity is preserved following solubilization is unknown. This study explored the tissue-specific bioactivity of soluble decellularized tendon and cartilage ECMs on human bone marrow-derived mesenchymal stem cells (MSCs) presented across different culture microenvironments, including two-dimensional (2D) tissue culture plastic, aligned electrospun nanofibers, cell pellets, and cell-seeded photocrosslinkable hydrogels. Tendon and cartilage ECMs were decellularized using established methods and solubilized either via pepsin digestion or urea extraction. The effect of soluble ECMs on cell proliferation and differentiation was initially explored by supplementing basal medium of human MSCs cultured on 2D tissue culture plastic. In subsequent experiments, MSCs were cultured on aligned electrospun nanofibers, ascell pellets, or encapsulated within photocrosslinkable methacrylated gelatin (GelMA) hydrogels. Urea-extracted tendon and cartilage ECMs were added as supplements. Pepsin-digested ECMs did not promote homologous differentiation in human MSCs, whether provided as a medium supplement or three-dimensional (3D) hydrogels. In contrast, urea-extracted ECMs tended to promote tissue-specific differentiation of MSCs cultured in 2D and 3D microenvironments. The application of the small molecule TGF-β signaling inhibitor SB-431542 largely negated the tissue-specific gene expression patterns mediated by tendon and cartilage ECMs. This suggests that the action of endogenous TGF-β was required, but was not sufficient, to impart tissue-specific bioactivity of urea-extracted ECMs. When urea-extracted cartilage ECM was incorporated within a photocurable GelMA hydrogel it independently enhanced chondrogenesis in encapsulated MSCs, and showed additive prochondrogenesis upon TGF-β supplementation in the medium. Urea-extracted ECM fractions of decellularized tendon and cartilage are soluble supplements capable of enhancing tissue-specific differentiation of adult stem cells. The online version of this article (doi:10.1186/s13287-017-0580-8) contains supplementary material, which is available to authorized users.