Journal: Advanced Healthcare Materials

Article Title: Repurposing FDA‐Approved Drugs for Temozolomide‐Resistant IDH1 Mutant Glioma Using High‐Throughput Miniaturized Screening on Droplet Microarray Chip

doi: 10.1002/adhm.202300591

Figure Lengend Snippet: Workflow for high‐throughput drug screening and hit validation. The droplet microarray chip uses minimal reagents (200 nL medium, 300 cells per spot) to screen 2208 FDA‐approved drugs rapidly and economically. The assay is validated using MTT assays in a standard 96‐well plate. On‐ and off‐target effects are assessed using Western blot and mass spectrometry. Drug target analysis is correlated with patient survival data from the Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA) databases to facilitate personalized clinical treatment.

Article Snippet: The droplet microarray (DMA) slides were purchased from Aquarray GmbH (catalog, IR0050241542, Eggenstein‐Leopoldshafen, Germany).

Techniques: High Throughput Screening Assay, Drug discovery, Biomarker Discovery, Microarray, Western Blot, Mass Spectrometry

Journal: Advanced Healthcare Materials

Article Title: Droplet Microarray Based Screening Identifies Proteins for Maintaining Pluripotency of hiPSCs

doi: 10.1002/adhm.202200718

Figure Lengend Snippet: Validation of the screening protocol. A) Representative confocal laser scanning microscope (CLSM) images of hiPSCs cultivated in 200 nL droplets on DMA slides coated with Matrigel and stained for six pluripotency markers: SOX2, OCT‐4A, NANOG, TRA‐1‐60, SSEA4, and TRA‐1‐81. Three independent experiments ( n = 3) obtained comparable results. DAPI was used to counterstain nuclei. Scale bars: 50 µm. B) Bright‐field images of hiPSCs cultivated on positive (Matrigel, MG + ) and negative (no Matrigel, MG − ) controls. Scale bar: 100 µm. C) NANOG expression level was used as a read out for the primary screening. IF staining was carried out on DMA followed by automated microscopy. All colonies in one image were from the single droplet. Scale bar: 50 µm. D) Quantification of mean brightness (mean fluorescence intensity) of hiPSCs cultured on positive and negative controls and stained for the pluripotency marker NANOG. The mean fluorescence intensity was calculated as the total fluorescence intensity divided by the area (in pixels). *** p < 0.001, significant differences positive control and negative controls. ( Z ʹ value is between 0.5 and 1, corresponding to a high‐quality screening assay). Data were presented as mean ± 3SD. ( n = 3) E) Representative CLSM images of E‐cadherin expression in hiPSCs cultured on positive and negative control coatings. E‐cadherin mediates cell–cell interactions and contributes to stem cell colony formation and pluripotency. Thus, on the positive control coating, undifferentiated hiPSCs show high expression of E‐cadherin, while differentiated hiPSCs on the negative control coating show low‐to‐no E‐cadherin expression. DAPI was used to counterstain nuclei. Scale bar: 25 µm.

Article Snippet: DMA slides with 1 mm side length square spots (catalogue number: G‐np‐102) were purchased from Aquarray GmbH (Eggenstein‐Leopoldshafen, Germany).

Techniques: Biomarker Discovery, Laser-Scanning Microscopy, Staining, Expressing, Microscopy, Fluorescence, Cell Culture, Marker, Positive Control, Screening Assay, Negative Control