Dynamic Click Hydrogels for Xeno-Free Culture of Induced Pluripotent Stem Cells.
Dynamic Click Hydrogels for Xeno-Free Culture of Induced Pluripotent Stem Cells.
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DOI：
10.1002/adbi.202000129
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发表时间：
2020-11
影响因子：
4.1
通讯作者：
Lin CC
中科院分区：
文献类型：
--
作者：
Arkenberg MR;Dimmitt NH;Johnson HC;Koehler KR;Lin CC
无xeno和化学定义的聚乙二醇（PEG）基水凝胶越来越多地用于诱导多能干细胞（iPSCs）的体外培养和分化。这些合成基质提供了高度可调的凝胶动力学和适应性材料特性，这对指导干细胞命运过程至关重要。在这项工作中，我们整合了顺序降冰片烯点击化学来形成合成的和动态可调的peg肽水凝胶，以促进人类iPSCs的培养和分化。具体来说，iPSCs被光封装在无xeno-降冰片烯水凝胶中，水凝胶由多臂peg -降冰片烯（PEG-NB）和蛋白酶不稳定的肽交联剂交联。这些基质被用来评估在基质可降解性、粘附配体、细胞密度和ROCK抑制作用的影响下iPSCs的生长。我们进一步采用四氮-降冰片烯（Tz-NB）逆电子需求Diels-Alder （iEDDA）反应来动态硬化这些载细胞的水凝胶。快速反应的Tz及其更稳定的衍生物甲基四嗪（mTz）被拴在多臂PEG上，生成单官能化的PEG-Tz和PEG-mTz，以及双官能化的PEG-Tz/mTz，它们很容易与PEG- nb反应，在载细胞的水凝胶中形成额外的交联。这种硬化方法的多功能性通过不同的tz修饰的大分子（例如，透明质酸- tz和肝素- tz）或通过PEG-Tz的间歇孵育来暂时调节硬化。最后，研究了z- nb介导的动态硬化对人多能干细胞的4D培养和内胚层分化的影响。总的来说，这种动态水凝胶平台提供了水凝胶交联的精细控制，作为无xeno和动态干细胞生态位。
Xeno-free and chemically-defined poly(ethylene glycol) (PEG)-based hydrogels are increasingly used for in vitro culture and differentiation of induced pluripotent stem cells (iPSCs). These synthetic matrices provide highly tunable gelation kinetics and adaptable material properties that are crucial for guiding stem cell fate processes. In this work, we integrated sequential norbornene-click chemistries for forming synthetic and dynamically tunable PEG-peptide hydrogels to promote human iPSCs culture and differentiation. Specifically, iPSCs were photo-encapsulated in xeno-free thiol-norbornene hydrogels crosslinked by multi-arm PEG-norbornene (PEG-NB) and protease-labile peptide crosslinkers. These matrices were used to evaluate growth of iPSCs under the influence of matrix degradability, adhesion ligands, cell density, and ROCK inhibition. We further employed tetrazine-norbornene (Tz-NB) inverse electron demand Diels-Alder (iEDDA) reaction to dynamically stiffen these cell-laden hydrogels. Fast reactive Tz and its more stable derivative methyltetrazine (mTz) were tethered to multi-arm PEG, yielding mono-functionalized PEG-Tz and PEG-mTz, as well as dual-functionalized PEG-Tz/mTz that readily reacted with PEG-NB to form additional crosslinks in the cell-laden hydrogels. The versatility of this stiffening approach was demonstrated with different Tz-modified macromers (e.g., hyaluronic acid-Tz and heparin-Tz) or by intermittent incubation of PEG-Tz for temporally regulated stiffening. Finally, the Tz-NB-mediated dynamic stiffening was explored for 4D culture and definitive endoderm differentiation of human iPSCs. Overall, this dynamic hydrogel platform affords exquisite controls of hydrogel crosslinking for serving as a xeno-free and dynamic stem cell niche.
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影响因子：
6.2
作者：
Cambria E;Renggli K;Ahrens CC;Cook CD;Kroll C;Krueger AT;Imperiali B;Griffith LG
通讯作者：
Griffith LG
影响因子：
64.8
作者：
Barriga EH;Franze K;Charras G;Mayor R
通讯作者：
Mayor R
影响因子：
4.6
作者：
Holt SE;Rakoski A;Jivan F;Pérez LM;Alge DL
通讯作者：
Alge DL
影响因子：
9.5
作者：
Hao Y;Song J;Ravikrishnan A;Dicker KT;Fowler EW;Zerdoum AB;Li Y;Zhang H;Rajasekaran AK;Fox JM;Jia X
通讯作者：
Jia X
影响因子：
14
作者：
Kerscher P;Turnbull IC;Hodge AJ;Kim J;Seliktar D;Easley CJ;Costa KD;Lipke EA
通讯作者：
Lipke EA