Droplet Microarray Based Screening Identifies Proteins for Maintaining Pluripotency of hiPSCs. Issue 18 (20th July 2022)

• Record Type: Journal Article
• Authors: Liu, Yanxi; Bertels, Sarah; Reischl, Markus; Peravali, Ravindra; Bastmeyer, Martin; Popova, Anna A.; Levkin, Pavel A.
• Is Part Of: Advanced healthcare materials. Volume 11:Issue 18(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-07-20
• Subjects: droplet microarrays -- high‐throughput screening -- hiPSCs -- pluripotency -- xeno‐free; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1002/adhm.202200718
• Languages: English
• ISSNs: 2192-2640

Abstract

Abstract: Human induced pluripotent stem cells (hiPSCs) are crucial for disease modeling, drug discovery, and personalized medicine. Animal‐derived materials hinderapplications of hiPSCs in medical fields. Thus, novel and well‐defined substrate coatings capable of maintaining hiPSC pluripotency are important for advancing biomedical applications of hiPSCs. Here a miniaturized droplet microarray (DMA) platform to investigate 11 well‐defined proteins, their 55 binary and 165 ternary combinations for their ability to maintainpluripotency of hiPSCs when applied as a surface coating, is used. Using this screening approach, ten protein group coatings are identified, which promote significantly higher NANOG expression of hiPSCs in comparison with Matrigel coating. With two of the identified coatings, long‐term pluripotency maintenance of hiPSCs and subsequent differentiation into three germ layers are achieved. Compared with conventional high‐throughput screening (HTS) in 96‐well plates, the DMA platform uses only 83 µL of protein solution (0.83 µg total protein) and only ≈2.8 × 10 5 cells, decreasing the amount of proteins and cells ≈860 and 25‐fold, respectively. The identified proteins will be essential for research and applications using hiPSCs, while the DMA platform demonstrates great potential for miniaturized HTS of scarce cells or expensive materials such as recombinant proteins. Abstract : A high‐throughput screening (HTS) of 231 well‐defined protein combinations, including 11 proteins, their 55 binary and 165 ternary combinations, using the miniaturized high‐density droplet microarray platform is performed. The newly discovered protein combinations hold great potential for both fundamental and applied research using hiPSCs. Droplet microarray platform will be essential for high‐throughput studies of expensive but important molecules.