Centre for Translational Bone, Joint and Soft Tissue Research, Faculty of Medicine and University Hospital Carl Gustav Carus, TU Dresden, Dresden, Germany.
Sci Rep. 2021 Mar 4;11(1):5130. doi: 10.1038/s41598-021-84384-6.
With the aim of understanding and recapitulating cellular interactions of hepatocytes in their physiological microenvironment and to generate an artificial 3D in vitro model, a co-culture system using 3D extrusion bioprinting was developed. A bioink based on alginate and methylcellulose (algMC) was first shown to be suitable for bioprinting of hepatocytes; the addition of Matrigel to algMC enhanced proliferation and morphology of them in monophasic scaffolds. Towards a more complex system that allows studying cellular interactions, we applied core-shell bioprinting to establish tailored 3D co-culture models for hepatocytes. The bioinks were specifically functionalized with natural matrix components (based on human plasma, fibrin or Matrigel) and used to co-print fibroblasts and hepatocytes in a spatially defined, coaxial manner. Fibroblasts acted as supportive cells for co-cultured hepatocytes, stimulating the expression of certain biomarkers of hepatocytes like albumin. Furthermore, matrix functionalization positively influenced both cell types in their respective compartments by enhancing their adhesion, viability, proliferation and function. In conclusion, we established a functional co-culture model with independently tunable compartments for different cell types via core-shell bioprinting. This provides the basis for more complex in vitro models allowing co-cultivation of hepatocytes with other liver-specific cell types to closely resemble the liver microenvironment.
为了理解和再现肝细胞在生理微环境中的细胞相互作用，并生成人工 3D 体外模型，开发了一种使用 3D 挤出式生物打印的共培养系统。首先，基于海藻酸盐和甲基纤维素（algMC）的生物墨水被证明适合肝细胞的生物打印；algMC 中添加 Matrigel 可增强它们在单相支架中的增殖和形态。为了建立更复杂的系统，以研究细胞相互作用，我们应用核壳生物打印技术来建立用于肝细胞的定制 3D 共培养模型。生物墨水专门用天然基质成分（基于人血浆、纤维蛋白或 Matrigel）进行功能化，并用于以空间定义的同轴方式共打印成纤维细胞和肝细胞。成纤维细胞作为共培养的肝细胞的支持细胞，刺激了白蛋白等肝细胞特定生物标志物的表达。此外，基质功能化通过增强细胞在各自隔室中的粘附、活力、增殖和功能，对两种细胞类型都产生了积极的影响。总之，我们通过核壳生物打印技术建立了一种具有独立可调隔室的功能性共培养模型，用于不同类型的细胞。这为更复杂的体外模型提供了基础，允许肝细胞与其他肝脏特异性细胞类型共培养，以更接近肝脏微环境。
