3D printing of sacrificial thioester elastomers using digital light processing for templating 3D organoid structures in soft biomatrices.
3D printing of sacrificial thioester elastomers using digital light processing for templating 3D organoid structures in soft biomatrices.
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DOI：
10.1088/1758-5090/ac1c98
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发表时间：
2021-09-02
期刊：
影响因子：
9
通讯作者：
Anseth KS
中科院分区：
文献类型：
--
作者：
Carberry BJ;Hergert JE;Yavitt FM;Hernandez JJ;Speckl KF;Bowman CN;McLeod RR;Anseth KS
生物织物允许在与细胞相关的尺寸尺度上对材料中的结构特征进行模板化，从而能够生成具有受控形式和功能的组织样结构。这对于生长类器官特别相关，其中生物化学和生物力学刺激的应用可用于引导干细胞的组装和分化，并形成类似于母体组织或器官的结构。最近，烧蚀激光扫描技术被用于在10-100 μm尺寸尺度的胶原蛋白水凝胶中创建3D悬垂特征，并支持肠类器官中的隐窝-绒毛结构。作为一种补充方法，为高通量图案化提供优势，我们使用数字光处理（DLP）打印硫酯官能化的聚（乙二醇）（PEG）弹性体，并产生可以模制到软（G' < 1000 Pa）水凝胶基底中的牺牲3D形状。具体地，将含有内部硫酯基团的3臂1.3kDa PEG硫醇和3臂1.6kDa PEG双烯光聚合以产生可降解弹性体。当在300 mM 2-巯基乙醇（pH 9.0）溶液中孵育时，1 mm厚、10 mm直径的弹性体圆盘在<2 h内降解。使用DLP，以分钟的数量级打印了具有37 ± 4 μm的临界尺寸、22 ± 5 μm的分辨率和小至50 μm的悬垂结构的特征阵列。将这些具有生理学相关特征的牺牲硫酯模具浇铸成型为基质胶，随后降解以产生具有高保真度的图案化空隙空间。在图案化的Matrigel基质上培养的肠干细胞形成符合基本图案的汇合单层。DLP打印的牺牲硫酯弹性体构建体提供了一种在软组织培养基质中制造3D类器官尺寸特征阵列的稳健且快速的方法，并且应该能够研究上皮几何形状和间距对肠干细胞（ISC）生长和分化的影响。
Biofabrication allows for the templating of structural features in materials on cellularly-relevant size scales, enabling the generation of tissue-like structures with controlled form and function. This is particularly relevant for growing organoids, where the application of biochemical and biomechanical stimuli can be used to guide the assembly and differentiation of stem cells and form architectures similar to the parent tissue or organ. Recently, ablative laser-scanning techniques was used to create 3D overhang features in collagen hydrogels at size scales of 10–100 μm and supported the crypt-villus architecture in intestinal organoids. As a complementary method, providing advantages for high-throughput patterning, we printed thioester functionalized poly(ethylene glycol) (PEG) elastomers using digital light processing (DLP) and created sacrificial, 3D shapes that could be molded into soft (G’ < 1000 Pa) hydrogel substrates. Specifically, 3-arm 1.3 kDa PEG thiol and 3-arm 1.6 kDa PEG norbornene, containing internal thioester groups, were photopolymerized to yield degradable elastomers. When incubated in a solution of 300 mM 2-mercaptoethanol (pH 9.0), 1 mm thick 10 mm diameter elastomer discs degraded in <2 h. Using DLP, arrays of features with critical dimensions of 37 ± 4 μm, resolutions of 22 ± 5 μm, and overhang structures as small as 50 μm, were printed on the order of minutes. These sacrificial thioester molds with physiologically relevant features were cast-molded into Matrigel and subsequently degraded to create patterned void spaces with high fidelity. Intestinal stem cells cultured on the patterned Matrigel matrices formed confluent monolayers that conformed to the underlying pattern. DLP printed sacrificial thioester elastomer constructs provide a robust and rapid method to fabricate arrays of 3D organoid-sized features in soft tissue culture substrates and should enable investigations into the effect of epithelial geometry and spacing on the growth and differentiation of intestinal stem cells (ISCs).
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DOI：
10.1038/s41578-021-00279-y
发表时间：
2021
期刊：
Nature reviews. Materials
影响因子：
--
作者：
Hofer M;Lutolf MP
通讯作者：
Lutolf MP
影响因子：
16.6
作者：
Chaudhuri O;Gu L;Darnell M;Klumpers D;Bencherif SA;Weaver JC;Huebsch N;Mooney DJ
通讯作者：
Mooney DJ
影响因子：
10
作者：
Hushka, Ella A.;Yavitt, F. Max;Anseth, Kristi S.
通讯作者：
Anseth, Kristi S.
影响因子：
29.4
作者：
Fairbanks, Benjamin D;Schwartz, Michael P;Halevi, Alexandra E;Nuttelman, Charles R;Bowman, Christopher N;Anseth, Kristi S
通讯作者：
Anseth, Kristi S
影响因子：
11
作者：
Higgins CI;Brown TE;Killgore JP
通讯作者：
Killgore JP