Rapid Bioorthogonal Chemistry Enables in Situ Modulation of the Stem Cell Behavior in 3D without External Triggers.
Rapid Bioorthogonal Chemistry Enables in Situ Modulation of the Stem Cell Behavior in 3D without External Triggers.
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DOI：
10.1021/acsami.8b07632
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发表时间：
2018-08-08
影响因子：
9.5
通讯作者：
Jia X
中科院分区：
文献类型：
--
作者：
Hao Y;Song J;Ravikrishnan A;Dicker KT;Fowler EW;Zerdoum AB;Li Y;Zhang H;Rajasekaran AK;Fox JM;Jia X
活细胞周围工程微环境的化学修饰代表了通过细胞-基质相互作用指导细胞行为的手段。本文提出了一种在活细胞培养期间采用快速的生物正交四嗪与反式环辛烯（TCO）亲二烯体连接来调节仿生合成细胞外基质（ECM）的性质的时间控制方法。这种方法是扩散控制的，细胞相容的，不依赖于光，催化剂或其他外部触发器。人骨髓间充质干细胞（hMSCs）最初包埋在水凝胶制备使用透明质酸携带巯基（HA-SH）和亲水性聚合物轴承丙烯酸酯和四嗪基团（POM-AT）。包括基质金属蛋白酶（MMP）可降解的肽交联剂使hMSC介导的重塑的合成环境。所得网络显示悬挂的四嗪基团，用于随后与TCO衍生物缀合。两天后，通过将携带多个拷贝的TCO的化学修饰的HA（HA-TCO）添加到围绕载有细胞的凝胶构建体的hMSC生长培养基中来增加基质的刚度。作为响应，细胞在细胞体周围放射状地形成小突起，而整体形状没有显著改变。相比之下，用TCO标记的细胞粘附基序修饰3D基质导致驻留细胞经历显著的肌动蛋白聚合，从圆形形状变为具有长细胞突起的纺锤体形态。在生长培养基中再培养7天后，定量分析显示，在mRNA水平上，RGD标记上调MMP 1的细胞表达，但下调I/III型胶原和腱生蛋白C的表达。然而，RGD标记不足以诱导经典的成骨细胞、成软骨细胞、成脂肪细胞或成纤维细胞/成肌纤维细胞分化。模块化方法允许容易地操纵合成ECM以调节细胞行为，因此可能适用于功能组织或组织模型的工程化。
Chemical modification of engineered microenvironments surrounding living cells represents a means for directing cellular behaviors through cell-matrix interactions. Presented here is a temporally controlled method for modulating the properties of biomimetic, synthetic extracellular matrices (ECM) during live cell culture employing the rapid, bioorthogonal tetrazine ligation with trans-cyclooctene (TCO) dienophiles. This approach is diffusion-controlled, cytocompatible and does not rely on light, catalysts or other external triggers. Human bone-marrow-derived mesenchymal stem cells (hMSCs) were initially entrapped in a hydrogel prepared using hyaluronic acid carrying sulfhydryl groups (HA-SH) and a hydrophilic polymer bearing both acrylate and tetrazine groups (POM-AT). Inclusion of a matrix metalloprotease (MMP)-degradable peptidic crosslinker enabled hMSC-mediated remodeling of the synthetic environment. The resultant network displayed dangling tetrazine groups for subsequent conjugation with TCO derivatives. Two days later, the stiffness of the matrix was increased by adding chemically modified HA carrying multiple copies of TCO (HA-TCO) to the hMSC growth media surrounding the cell-laden gel construct. In response, cells developed small processes radially around the cell body without a significant alteration of the overall shape. By contrast, modification of the 3D matrix with a TCO-tagged cell-adhesive motif caused the resident cells to undergo significant actin polymerization, changing from a rounded shape to spindle morphology with long cellular processes. After additional 7 days of culture in the growth media, quantitative analysis showed that, at the mRNA level, RGD tagging upregulated cellular expression of MMP1, but downregulated the expression of collagen I/III and tenascin C. RGD tagging, however, was not sufficient to induce the classic osteoblastic, chondrogenic, adipogenic, or fibroblastic/myofibroblastic differentiation. The modular approach allows facile manipulation of synthetic ECM to modulate cell behavior, thus potentially applicable to the engineering of functional tissues or tissue models.
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影响因子：
15
作者：
Blackman, Melissa L.;Royzen, Maksim;Fox, Joseph M.
通讯作者：
Fox, Joseph M.
影响因子：
2.8
作者：
Gong, Yong-Hua;Miomandre, Fabien;Clavier, Gilles
通讯作者：
Clavier, Gilles
影响因子：
9.7
作者：
Dicker, Kevin T.;Gurski, Lisa A.;Pradhan-Bhatt, Swati;Witt, Robert L.;Farach-Carson, Mary C.;Jia, Xinqiao
通讯作者：
Jia, Xinqiao
影响因子：
41.2
作者：
Chaudhuri O;Gu L;Klumpers D;Darnell M;Bencherif SA;Weaver JC;Huebsch N;Lee HP;Lippens E;Duda GN;Mooney DJ
通讯作者：
Mooney DJ
影响因子：
64.5
作者：
DISTEL, RJ;RO, HS;SPIEGELMAN, BM
通讯作者：
SPIEGELMAN, BM