Shrestha Sunil, Vanga Manav Goud, Jonnadula Charishma, Acharya Prabha, Lee Minseong, Lee Moo-Yeal
Department of Biomedical Engineering, University of North Texas, Denton, TX, USA.
Bioprinting Laboratories Inc., Dallas, TX, USA.
Methods Mol Biol. 2025 Jan 17. doi: 10.1007/7651_2024_603.
Human liver organoids (HLOs) derived from pluripotent stem cells hold potential for disease modeling and high-throughput compound screening due to their architectural and functional resemblance to human liver tissues. However, reproducible, scale-up production of HLOs for high-throughput screening (HTS) presents challenges. These include the high costs of additives and growth factors required for cell differentiation, variability in organoid size and function from batch to batch, suboptimal maturity of HLOs compared to primary hepatocytes, and low assay throughput due to excessive manual processes and the absence of assay-ready plates with HLOs. To address some of these issues, here we present standard operating procedures (SOPs) for the scale-up production of HLOs using a pillar plate through microarray 3D bioprinting. This technology facilitates the rapid, uniform seeding of foregut cells onto the pillar plate, maintaining cell viability and enabling the scale-up generation of HLOs. The assay-ready pillar plate with HLOs is suitable for compound testing, as well as in situ organoid staining and analysis.
源自多能干细胞的人肝脏类器官(HLOs)因其在结构和功能上与人类肝脏组织相似,在疾病建模和高通量化合物筛选方面具有潜力。然而,为高通量筛选(HTS)可重复地扩大HLOs的生产面临挑战。这些挑战包括细胞分化所需添加剂和生长因子的高成本、不同批次类器官大小和功能的差异、与原代肝细胞相比HLOs成熟度欠佳,以及由于过多的手工操作和缺乏带有HLOs的即用型检测板导致的低检测通量。为了解决其中一些问题,我们在此展示使用柱板通过微阵列3D生物打印扩大HLOs生产的标准操作程序(SOPs)。该技术有助于将前肠细胞快速、均匀地接种到柱板上,维持细胞活力并实现HLOs的扩大生产。带有HLOs的即用型柱板适用于化合物测试,以及类器官原位染色和分析。
ACS Biomater Sci Eng. 2025-1-13
Adv Healthc Mater. 2024-8