Department of Medical Engineering, University of South Florida, Tampa, FL, USA.
Department of Electrical & Computer Engineering, Old Dominion University, Norfolk, VA, USA.
Bioelectrochemistry. 2021 Aug;140:107827. doi: 10.1016/j.bioelechem.2021.107827. Epub 2021 Apr 26.
We developed and characterized a 3D collagen hydrogel model for B16.F10 melanoma tumors. Cells in this 3D environment exhibited lower proliferation than cells in the conventional 2D culture environment. Interestingly, the basal expression levels of several genes varied when compared to conventionally grown cells. In each growth environment, a significant number of melanoma cells were transfected by plasmid electroporation (electrotransfer), although expression could only be ascertained on the surface of the 3D constructs. Cellular responses to plasmid entry as demonstrated by pro-inflammatory cytokine and chemokine upregulation varied based on the growth environment, as did the mRNA levels of several putative DNA-specific pattern recognition receptors (DNA sensors). Unexpectedly, when plasmid DNA was delivered while cells where attached in the 2D or 3D environments, the mRNAs of the DNA sensor p204 and the inflammatory mediator TNFα were regulated in cells receiving pulses only. However, we were unable to confirm coordinate upregulation of TNFα and p204 proteins. This study confirms that cell responses differ significantly based on their environment, and demonstrates the difficulty of extending experimental observations between cell environments.
我们开发并表征了一种用于 B16.F10 黑色素瘤肿瘤的 3D 胶原水凝胶模型。与传统的 2D 培养环境相比，该 3D 环境中的细胞增殖率较低。有趣的是，与常规生长的细胞相比，几种基因的基础表达水平存在差异。在每种生长环境中，通过质粒电穿孔（电转移）转染了大量黑色素瘤细胞，尽管仅能在 3D 构建体的表面确定表达。细胞对质粒进入的反应表现为促炎细胞因子和趋化因子的上调，这取决于生长环境，几种潜在的 DNA 特异性模式识别受体（DNA 传感器）的 mRNA 水平也存在差异。出乎意料的是，当在 2D 或 3D 环境中附着的细胞递送质粒 DNA 时，仅在接受脉冲的细胞中调节 DNA 传感器 p204 和炎症介质 TNFα 的 mRNA。然而，我们无法确认 TNFα 和 p204 蛋白的协调上调。这项研究证实，细胞反应根据其环境有很大差异，并证明了在细胞环境之间扩展实验观察结果的困难。
