Efficient One-Step Production of Microencapsulated Hepatocyte Spheroids with Enhanced Functions.
Efficient One-Step Production of Microencapsulated Hepatocyte Spheroids with Enhanced Functions.
复制标题
DOI：
10.1002/smll.201502932
复制
发表时间：
2016-05
期刊：
影响因子：
--
通讯作者：
Leong KW
中科院分区：
文献类型：
--
作者：
Chan HF;Zhang Y;Leong KW
在水凝胶中微囊化的肝细胞球体可以在各种能力上为肝脏研究做出贡献。传统的微胶囊化方法会在每个微凝胶中产生可变数量的球体，并且需要将球体装入凝胶中。本文报道了一种微流控技术，可以绕过球体加载的步骤，控制球体特性。双乳（DE）微滴在4小时内生成微囊化的同型或异型肝细胞球体（均为直径<200 μm的单个球体），其功能增强。当用海藻酸盐-胶原复合水凝胶代替海藻酸盐时，培养24天的肝细胞功能（白蛋白分泌、尿素分泌和细胞色素P450活性）得到改善，微凝胶的组成是可调的。在海藻酸-胶原中培养的肝细胞球体也比在胶原-三明治结构中培养的肝细胞表现更好。此外，当肝细胞与内皮祖细胞（作为一种新的支持细胞来源）以5比1的最佳比例共同培养形成复合球体时，肝细胞功能显著增强，海藻酸胶原包埋可进一步增强复合球体的功能。这种具有高产量、多功能性和均匀性的微封装球体形成技术被设想为肝组织工程和生物制造的使能技术。
Hepatocyte spheroids microencapsulated in hydrogels can contribute to liver research in various capacities. Conventional approach of microencapsulating spheroids produces variable number of spheroids per microgel and requires an extra step of spheroid loading into the gel. Here, we report a microfluidics technology bypassing the step of spheroid loading and controlling the spheroid characteristics. Double-emulsion (DE) droplets were used to generate microencapsulated homotypic or heterotypic hepatocyte spheroids (all as single spheroids <200 μm in diameter) with enhanced functions in 4 hours. The composition of the microgel is tunable as demonstrated by improved hepatocyte functions during 24 day culture (albumin secretion, urea secretion and cytochrome P450 activity) when alginate-collagen composite hydrogel was used instead of alginate. Hepatocyte spheroids in alginate-collagen also performed better than hepatocytes cultured in collagen-sandwich configuration. Moreover, hepatocyte functions were significantly enhanced when hepatocytes and endothelial progenitor cells (used as a novel supporting cell source) were co-cultured to form composite spheroids at an optimal ratio of 5 to 1, which could be further boosted when encapsulated in alginate-collagen. This microencapsulated-spheroid formation technology with high yield, versatility and uniformity is envisioned to be an enabling technology for liver tissue engineering as well as biomanufacturing.