The Effect of Thiol Structure on Allyl Sulfide Photodegradable Hydrogels and their Application as a Degradable Scaffold for Organoid Passaging.
The Effect of Thiol Structure on Allyl Sulfide Photodegradable Hydrogels and their Application as a Degradable Scaffold for Organoid Passaging.
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DOI：
10.1002/adma.201905366
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发表时间：
2020-07
期刊：
影响因子：
--
通讯作者：
Anseth KS
中科院分区：
文献类型：
--
作者：
Yavitt FM;Brown TE;Hushka EA;Brown ME;Gjorevski N;Dempsey PJ;Lutolf MP;Anseth KS
肠道类器官已被证明是许多旨在理解和治疗疾病的基础和转化研究的有用体外模型。然而，它们的常规培养依赖于动物源性基质，这限制了它们在临床应用中的效用。事实上，很少有完全确定的合成水凝胶系统能够支持肠道类器官的发育和扩增。在此，我们介绍一种烯丙基硫化物光降解水凝胶，它通过自由基加成 - 断裂链转移反应实现快速降解，以支持肠道类器官的常规传代。我们首先利用剪切流变学来表征可溶性硫醇和烯丙基硫化物交联结构对降解动力学的影响。在可溶性硫醇（15 mM谷胱甘肽）和光引发剂（1 mM苯基 - 2,4,6 - 三甲基苯甲酰基次膦酸锂）存在的情况下，用365nm光（5 mW/cm²）照射，可使水凝胶在不到15秒内完全降解。烯丙基硫化物水凝胶被用于支持单个肠道干细胞形成上皮集落，并且利用快速光降解实现干细胞集落的重复传代，而形态或类器官形成潜能没有损失。这个平台可用于支持肠道类器官的长期培养，有可能取代对动物源性基质的需求，同时还能针对感兴趣的类器官对水凝胶特性进行系统性调整。
Intestinal organoids have proven to be useful in vitro models for many basic and translational studies aimed at understanding and treating disease. However, their routine culture relies on animal derived matrices that limit their utility for clinical applications. In fact, there are few fully defined, synthetic hydrogel systems that allow for the development and expansion of intestinal organoids. Here, we present an allyl sulfide photodegradable hydrogel, achieving rapid degradation through radical addition-fragmentation chain transfer reactions, to support the routine passaging of intestinal organoids. We use shear rheology to first characterize the effect of soluble thiol and allyl sulfide crosslink structures on degradation kinetics. Irradiation with 365nm light (5 mW cm−2) in the presence of a soluble thiol (glutathione at 15 mM), and a photoinitiator (lithium phenyl-2,4,6-trimethylbenzoylphosphinate at 1mM), led to complete hydrogel degradation in less than 15 seconds. Allyl sulfide hydrogels were used to support the formation of epithelial colonies from single intestinal stem cells, and rapid photodegradation was used to achieve repetitive passaging of stem cell colonies without loss in morphology or organoid formation potential. This platform could be used to support long term culture of intestinal organoids, potentially replacing the need for animal derived matrices, while also allowing systematic variations to the hydrogel properties tailored for the organoid of interest.
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DOI：
10.1002/adma.201304847
发表时间：
2014-04-23
期刊：
Advanced materials (Deerfield Beach, Fla.)
影响因子：
--
作者：
Gandavarapu NR;Azagarsamy MA;Anseth KS
通讯作者：
Anseth KS
DOI：
10.1155/2012/248759
发表时间：
2012
期刊：
Journal of signal transduction
影响因子：
--
作者：
Benoit YD;Groulx JF;Gagné D;Beaulieu JF
通讯作者：
Beaulieu JF
影响因子：
4.7
作者：
Colak, Burcu;Da Silva, Julio C. S.;Gautrot, Julien E.
通讯作者：
Gautrot, Julien E.
影响因子：
5.5
作者：
Fairbanks BD;Singh SP;Bowman CN;Anseth KS
通讯作者：
Anseth KS
影响因子：
3.4
作者：
BIAGLOW, JE;VARNES, ME;EPP, ER
通讯作者：
EPP, ER