PEG-4MAL hydrogels for human organoid generation, culture, and in vivo delivery.
PEG-4MAL hydrogels for human organoid generation, culture, and in vivo delivery.
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DOI：
10.1038/s41596-018-0036-3
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发表时间：
2018-09
期刊：
影响因子：
14.8
通讯作者：
García AJ
中科院分区：
文献类型：
--
作者：
Cruz-Acuña R;Quirós M;Huang S;Siuda D;Spence JR;Nusrat A;García AJ
人多能干细胞（hPSC）衍生的类器官（HO）的体外分化促进了类似于天然人组织的多细胞三维结构的产生。目前大多数用于生成HO的方法依赖于基质胶，基质胶是由Engelbreth-Holm-Swarm小鼠肉瘤细胞分泌的一种定义不清的基底膜衍生物，限制了HO用于再生医学应用的潜在用途。在这里，我们描述了一个协议，用于合成一个完全定义的，合成水凝胶，支持生成和文化的HO。模块化的细胞包封水凝胶由四臂聚（乙二醇）大分子单体形成，该大分子单体在每个末端具有马来酰亚胺基团（PEG-4 MAL），并与含半胱氨酸的粘附肽缀合，并通过蛋白酶可降解肽交联。该方案还包括用于将PEG-4 MAL水凝胶包封的HO在体内局部递送至损伤的小鼠结肠的指南。PEG-4 MAL水凝胶支持HO的植入并加速结肠伤口修复。因此，这种培养和递送策略可用于开发基于HO的疗法以治疗损伤和疾病。水凝胶和组织制备以及随后的包封可在2.5-3.5小时内进行。一旦HO在合成水凝胶中培养至少14天，它们就可以在5小时内制备并递送到小鼠结肠。
In vitro differentiation of human pluripotent stem cell (hPSC)-derived organoids (HOs) facilitates the production of multicellular three-dimensional structures analogous to native human tissues. Most current methods for the generation of HOs rely on Matrigel, a poorly defined basement membrane derivative secreted by Engelbreth-Holm-Swarm mouse sarcoma cells, limiting the potential use of HOs for regenerative medicine applications. Here, we describe a protocol for the synthesis of a fully defined, synthetic hydrogel that supports the generation and culture of HOs. Modular, cell-encapsulating hydrogels are formed from a four-armed poly(ethylene glycol) macromer that has maleimide groups at each terminus (PEG-4MAL) and is conjugated to cysteine-containing adhesive peptides and cross-linked via protease-degradable peptides. The protocol also includes guidelines for the localized in vivo delivery of PEG-4MAL hydrogel-encapsulated HOs to injured mouse colon. The PEG-4MAL hydrogel supports the engraftment of the HOs and accelerates colonic wound repair. This culture and delivery strategy can thus be used to develop HO-based therapies to treat injury and disease. Hydrogel and tissue preparation and subsequent encapsulation can be performed within 2.5-3.5 h. Once HOs have been cultured in synthetic hydrogels for at least 14 d, they can be prepared and delivered to the mouse colon in under 5 h.
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影响因子：
14.8
作者：
通讯作者：
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影响因子：
4.6
作者：
Wells, James M.;Spence, Jason R.
通讯作者：
Spence, Jason R.
影响因子：
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作者：
Dye BR;Hill DR;Ferguson MA;Tsai YH;Nagy MS;Dyal R;Wells JM;Mayhew CN;Nattiv R;Klein OD;White ES;Deutsch GH;Spence JR
通讯作者：
Spence JR
影响因子：
1.2
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通讯作者：
Bettenworth, Dominik
影响因子：
14.8
作者：
Gjorevski, Nikolce;Lutolf, Matthias P.
通讯作者：
Lutolf, Matthias P.