Department of Biomedical Engineering, Purdue School of Engineering and Technology, Indiana University-Purdue University at Indianapolis, Indianapolis, IN 46202, USA.
Biomaterials. 2013 Jul;34(21):5117-27. doi: 10.1016/j.biomaterials.2013.03.086. Epub 2013 Apr 19.
A highly tunable synthetic biomimetic hydrogel platform was developed to study the growth and morphogenesis of pancreatic ductal epithelial cells (PDEC) under the influence of a myriad of instructive cues. A PDEC line, PANC-1, was used as a model system to illustrate the importance of matrix compositions on cell fate determination. PANC-1 is an immortalized ductal epithelial cell line widely used in the study of pancreatic tumor cell behaviors. PANC-1 cells are also increasingly explored as a potential cell source for endocrine differentiation. Thus far, most studies related to PANC-1, among other PDEC lines, are performed on 2D culture surfaces. Here, we evaluated the effect of matrix compositions on PANC-1 cell growth and morphogenesis in 3D. Specifically, PANC-1 cells were encapsulated in PEG-based hydrogels prepared by step-growth thiol-ene photopolymerization. It was found that thiol-ene hydrogels provided a cytocompatible environment for encapsulation and 3D culture of PANC-1 cells. In contrast to a monolayer morphology on 2D culture surfaces, PANC-1 cells formed clusters in 3D thiol-ene hydrogels within 4 days of culture. After culturing for 10 days, however, the growth and structures of these clusters were significantly impacted by gel matrix properties, including sensitivity of the matrix to proteases, stiffness of the matrix, and ECM-mimetic motifs. The use of matrix metalloproteinase (MMP) sensitive linker or the immobilization of fibronectin-derived RGDS ligand in the matrix promoted PANC-1 cell growth and encouraged them to adopt ductal cyst-like structures. On the other hand, the encapsulated cells formed smaller and more compact aggregates in non-MMP responsive gels. The incorporation of laminin-derived YIGSR peptide did not enhance cell growth and caused the cells to form compact aggregates. Immobilized YIGSR also enhanced the expression of epithelial cell markers including β-catenin and E-cadherin. These studies have established PEG-peptide hydrogels formed by thiol-ene photo-click reaction as a suitable platform for studying and manipulating pancreatic epithelial cell growth and morphogenesis in 3D.
开发了一种高度可调的仿生合成水凝胶平台，以在多种指导信号的影响下研究胰腺导管上皮细胞（PDEC）的生长和形态发生。使用 PANC-1 细胞系作为模型系统来说明基质组成对细胞命运决定的重要性。PANC-1 是一种广泛用于研究胰腺肿瘤细胞行为的永生化导管上皮细胞系。PANC-1 细胞也越来越多地被探索作为内分泌分化的潜在细胞来源。到目前为止，大多数与 PANC-1 相关的研究，以及其他 PDEC 系的研究，都是在二维培养表面上进行的。在这里，我们评估了基质组成对 PANC-1 细胞在 3D 中生长和形态发生的影响。具体来说，将 PANC-1 细胞包封在通过逐步增长硫醇-烯光聚合制备的基于 PEG 的水凝胶中。结果发现，硫醇-烯水凝胶为 PANC-1 细胞的包封和 3D 培养提供了细胞相容的环境。与二维培养表面上的单层形态相比，PANC-1 细胞在 3D 硫醇-烯水凝胶中培养 4 天后形成细胞团。然而，培养 10 天后，这些细胞团的生长和结构受到凝胶基质特性的显著影响，包括基质对蛋白酶的敏感性、基质的刚性和 ECM 模拟基序。使用基质金属蛋白酶（MMP）敏感连接子或在基质中固定纤连蛋白衍生的 RGDS 配体促进了 PANC-1 细胞的生长，并促使它们采用导管样囊肿结构。另一方面，在非 MMP 响应凝胶中，包封的细胞形成更小且更紧凑的聚集体。整合层粘连蛋白衍生的 YIGSR 肽并没有增强细胞生长并导致细胞形成紧密聚集体。固定的 YIGSR 还增强了上皮细胞标志物的表达，包括β-连环蛋白和 E-钙粘蛋白。这些研究建立了通过硫醇-烯光点击反应形成的 PEG-肽水凝胶作为研究和操纵胰腺上皮细胞在 3D 中生长和形态发生的合适平台。
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