Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA.
Department of Ophthalmology and Visual Neurosciences, University of Minnesota, Minneapolis, MN, USA.
SLAS Technol. 2021 Jun;26(3):287-299. doi: 10.1177/2472630320972110. Epub 2020 Dec 9.
Derivation and differentiation of human induced pluripotent stem cells (hiPSCs) provide the opportunity to generate medically important cell types from individual patients and patient populations for research and the development of potential cell therapies. This technology allows disease modeling and drug screening to be carried out using diverse population cohorts and with more relevant cell phenotypes than can be accommodated using traditional immortalized cell lines. However, technical complexities in the culture and differentiation of hiPSCs, including lack of scale and standardization and prolonged experimental timelines, limit the adoption of this technology for many large-scale studies, including personalized drug screening. The entry of reproducible end-to-end automated workflows for hiPSC culture and differentiation, demonstrated on commercially available platforms, provides enhanced accessibility of this technology for both research laboratories and commercial pharmaceutical testing. Here we have utilized TECAN Fluent automated cell culture workstations to perform hiPSC culture and differentiation in a reproducible and scalable process to generate patient-derived retinal pigment epithelial cells for downstream use, including drug testing. hiPSCs derived from multiple donors with age-related macular degeneration (AMD) were introduced into our automated workflow, and cell lines were cultured and differentiated into retinal pigment epithelium (RPE). Donor hiPSC-RPE lines were subsequently entered in an automated drug testing workflow to measure mitochondrial function after exposure to "mitoactive" compounds. This work demonstrates scalable, reproducible culture and differentiation of hiPSC lines from individuals on the TECAN Fluent platform and illustrates the potential for end-to-end automation of hiPSC-based personalized drug testing.
人诱导多能干细胞（hiPSCs）的诱导和分化为从个体患者和患者群体中生成对医学重要的细胞类型提供了机会，可用于研究和开发潜在的细胞疗法。该技术允许使用不同的人群队列进行疾病建模和药物筛选，并具有比传统永生化细胞系更相关的细胞表型。然而，hiPSCs 的培养和分化技术复杂，包括缺乏规模和标准化以及实验时间延长，限制了该技术在许多大规模研究中的应用，包括个性化药物筛选。可在商业平台上演示的可重复的端到端自动化 hiPSC 培养和分化工作流程的引入，为研究实验室和商业制药测试提供了更方便地使用这项技术的途径。在这里，我们利用 TECAN Fluent 自动化细胞培养工作站，以可重复和可扩展的方式进行 hiPSC 培养和分化，生成用于下游应用（包括药物测试）的患者来源的视网膜色素上皮细胞。将来自多位年龄相关性黄斑变性（AMD）患者的 hiPSC 引入我们的自动化工作流程，并培养和分化细胞系成为视网膜色素上皮（RPE）。随后，供体 hiPSC-RPE 系被纳入自动化药物测试工作流程，以测量暴露于“有丝分裂活性”化合物后的线粒体功能。这项工作证明了在 TECAN Fluent 平台上可从个体中进行可扩展、可重复的 hiPSC 培养和分化，并说明了基于 hiPSC 的个性化药物测试的端到端自动化的潜力。
