Integrating sensing hydrogel microstructures into micropatterned hepatocellular cocultures.
Integrating sensing hydrogel microstructures into micropatterned hepatocellular cocultures.
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DOI：
10.1021/la803635r
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发表时间：
2009-04-09
期刊：
影响因子：
--
通讯作者：
Revzin A
中科院分区：
文献类型：
--
作者：
Lee JY;Shah SS;Yan J;Howland MC;Parikh AN;Pan T;Revzin A
本文描述了一种微加工衍生的方法，用于定义不同细胞群之间的相互作用，并将生物传感器与细胞微图案相结合。在该方法中，采用光刻胶光刻法在硅烷改性的玻璃基底上对细胞粘附配体（胶原（I））进行微图案化。然后使用聚（乙二醇）（PEG）光刻法制造与现有胶原（I）结构域配准的水凝胶微结构。以这种方式改性的玻璃基底具有三种类型的微图案化区域：细胞粘附性胶原（I）结构域、适度粘附性硅烷化玻璃区域和非粘附性PEG水凝胶区域。将该基质与原代大鼠肝细胞或HepG2细胞一起孵育，导致肝细胞附着在胶原蛋白（I）结构域上，而在硅烷改性的玻璃区域或水凝胶结构域上未观察到粘附。将3T3成纤维细胞添加到相同表面上，附着在肝细胞周围的玻璃区域上，完成共培养。值得注意的是，PEG水凝胶微结构保持不含细胞，并用于将肝细胞与成纤维细胞“隔离”，从而限制了细胞类型之间的交流。我们还证明，水凝胶微结构内的酶分子的截留并不损害PEG的非污染特性。在此结果的基础上，将含有辣根过氧化物酶（HRP）的水凝胶微结构整合到微图案化的共培养物中，并用于检测培养基中的过氧化氢。这里描述的表面微图案化方法可能在未来用于同时定义和检测两种不同细胞类型之间的内分泌信号。
This paper describes a microfabrication-derived approach for defining interactions between distinct groups of cells and integrating biosensors with cellular micropatterns. In this approach, photoresist lithography was employed to micropattern cells adhesive ligand (collagen (I)) on silane-modified glass substrates. Poly(ethylene glycol) (PEG) photolithography was then used to fabricate hydrogel microstructures in registration with existing collagen (I) domains. A glass substrate modified in this manner had three types of micrpatterned regions: cell-adhesive collagen (I) domains, moderately adhesive silanized glass regions and non-adhesive PEG hydrogel regions. Incubation of this substrate with primary rat hepatocytes or HepG2 cells resulted in attachment of hepatic cells on collagen (I) domains with no adhesion observed on silane-modified glass regions or hydrogel domains. 3T3 fibroblasts added onto the same surface attached on the glass regions around the hepatocytes, completing the co-culture. Significantly, PEG hydrogel microstructures remained free of cells and were used to “fence” hepatocytes from fibroblasts, thus limiting communication between the cell types. We also demonstrated that entrapment of enzyme molecules inside hydrogel microstructures did not compromise non-fouling properties of PEG. Building on this result, horse radish peroxidase (HRP)-containing hydrogel microstructures were integrated into micropatterned co-cultures and were used to detect hydrogen peroxide in the culture medium. The surface micropatterning approach described here may be used in the future to simultaneously define and detect endocrine signaling between two distinct cell types.
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影响因子：
7.4
作者：
Seong, GH;Heo, J;Crooks, RM
通讯作者：
Crooks, RM
影响因子：
15
作者：
Jiang, XY;Ferrigno, R;Whitesides, GM
通讯作者：
Whitesides, GM
影响因子：
7.4
作者：
Russell, RJ;Pishko, MV;Wild, JR
通讯作者：
Wild, JR
DOI：
10.1073/pnas.0608660104
发表时间：
2007-04-03
影响因子：
11.1
作者：
Hui, Elliot E.;Bhatia, Sangeeta N.
通讯作者：
Bhatia, Sangeeta N.
影响因子：
6.1
作者：
Revzin, A;Sekine, K;Toner, M
通讯作者：
Toner, M