Nonlinear 3D projection printing of concave hydrogel microstructures for long-term multicellular spheroid and embryoid body culture.
Nonlinear 3D projection printing of concave hydrogel microstructures for long-term multicellular spheroid and embryoid body culture.
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DOI：
10.1039/c5lc00159e
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发表时间：
2015-06-07
期刊：
影响因子：
6.1
通讯作者：
Chen SC
中科院分区：
文献类型：
--
作者：
Hribar KC;Finlay D;Ma X;Qu X;Ondeck MG;Chung PH;Zanella F;Engler AJ;Sheikh F;Vuori K;Chen SC
单个多细胞球体和胚状体（EBs）的长期培养和监测仍然是体外细胞繁殖的挑战。在这里，我们使用了一种连续的3D投影打印方法-对非线性曝光进行了重要的修改-来生成凹面水凝胶微结构，该微结构允许球体生长和长期维护，而不需要球体转移。乳腺癌球状体生长到10天的凹结构显示缺氧核心和坏死的迹象，使用免疫荧光和组织化学染色，在体内的肿瘤微环境的关键特征。由在水凝胶上生长的诱导多能干细胞（iPSC）组成的EB在3 d时表现出窄的尺寸分布和未分化的标记，随后在10 d时通过存在空腔和三个胚层的染色而表现出分化的迹象。这些发现证明了一种用于长期（例如，在第2天超过球体形成，并进行培养基交换）3D细胞培养的新方法，该方法应该能够帮助癌症球体研究以及胚胎发生和iPSC EB患者来源的疾病建模。
Long-term culture and monitoring of individual multicellular spheroids and embryoid bodies (EBs) remains a challenge for in vitro cell propogation. Here, we used a continuous 3D projection printing approach – with an important modification of nonlinear exposure — to generate concave hydrogel microstructures that permit spheroid growth and long-term maintenance, without the need for spheroid transfer. Breast cancer spheroids grown to 10 d in the concave structures showed hypoxic cores and signs of necrosis using immunofluorescent and histochemical staining, key features of the tumor microenvironment in vivo. EBs consisting of induced pluripotent stem cells (iPSCs) grown on the hydrogels demonstrated narrow size distribution and undifferentiated markers at 3 d, followed by signs of differentiation by the presence of cavities and staining of the three germ layers at 10 d. These findings demonstrate a new method for long-term (e.g. beyond spheroid formation at day 2, and with media exchange) 3D cell culture that should be able to assist in cancer spheroid studies as well as embryogenesis and patient-derived disease modeling with iPSC EBs.
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影响因子：
3.7
作者：
Ungrin MD;Joshi C;Nica A;Bauwens C;Zandstra PW
通讯作者：
Zandstra PW
影响因子：
--
作者：
Thoma, Claudio R.;Stroebel, Simon;Kelm, Jens M.
通讯作者：
Kelm, Jens M.
DOI：
10.1016/j.jconrel.2012.04.045
发表时间：
2012-12-10
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Mehta G;Hsiao AY;Ingram M;Luker GD;Takayama S
通讯作者：
Takayama S
影响因子：
19
作者：
Huh, Dongeun;Hamilton, Geraldine A.;Ingber, Donald E.
通讯作者：
Ingber, Donald E.
影响因子：
--
作者：
Fukuda, J;Nakazawa, K
通讯作者：
Nakazawa, K