Wang Alex J, Allen Allysa, Sofman Marianna, Sphabmixay Pierre, Yildiz Ece, Griffith Linda G
Biological Engineering Department, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA.
Mechanical Engineering Department, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA; Whitehead Institute for Biomedical Research, 455 Main Street, Cambridge, MA, 02142, USA.
Adv Nanobiomed Res. 2022 Jan;2(1). doi: 10.1002/anbr.202100049. Epub 2021 Nov 19.
models of human liver functions are used across a diverse range of applications in preclinical drug development and disease modeling, with particular increasing interest in models that capture facets of liver inflammatory status. This study investigates how the interplay between biophysical and biochemical microenvironment cues influence phenotypic responses, including inflammation signatures, of primary human hepatocytes (PHH) cultured in a commercially available perfused bioreactor. A 3D printing-based alginate microwell system was designed to form thousands of hepatic spheroids in a scalable manner as a comparator 3D culture modality to the bioreactor. Soft, synthetic extracellular matrix (ECM) hydrogel scaffolds with biophysical properties mimicking features of liver were engineered to replace polystyrene scaffolds, and the biochemical microenvironment was modulated with a defined set of growth factors and signaling modulators. The supplemented media significantly increased tissue density, albumin secretion, and CYP3A4 activity but also upregulated inflammatory markers. Basal inflammatory markers were lower for cells maintained in ECM hydrogel scaffolds or spheroid formats than polystyrene scaffolds, while hydrogel scaffolds exhibited the most sensitive response to inflammation as assessed by multiplexed cytokine and RNA-seq analyses. Together, these engineered 3D liver microenvironments provide insights for probing human liver functions and inflammatory response .
人类肝脏功能模型在临床前药物开发和疾病建模的各种应用中得到了广泛应用，人们对能够捕捉肝脏炎症状态各个方面的模型的兴趣尤其日益浓厚。本研究调查了生物物理和生化微环境线索之间的相互作用如何影响在市售灌注生物反应器中培养的原代人肝细胞（PHH）的表型反应，包括炎症特征。设计了一种基于3D打印的藻酸盐微孔系统，以可扩展的方式形成数千个肝球体，作为生物反应器的一种比较性3D培养模式。设计了具有模仿肝脏特征的生物物理特性的柔软合成细胞外基质（ECM）水凝胶支架来替代聚苯乙烯支架，并使用一组特定的生长因子和信号调节剂来调节生化微环境。添加的培养基显著增加了组织密度、白蛋白分泌和CYP3A4活性，但也上调了炎症标志物。与聚苯乙烯支架相比，在ECM水凝胶支架或球体形式中培养的细胞的基础炎症标志物较低，而通过多重细胞因子和RNA测序分析评估，水凝胶支架对炎症表现出最敏感的反应。总之，这些工程化的3D肝脏微环境为探究人类肝脏功能和炎症反应提供了见解。
