Transcriptome profiling of myeloid cell lines in Gelatin Methacrylate Hydrogels
Transcriptome profiling of myeloid cell lines in Gelatin Methacrylate Hydrogels
Cell lines were analyzed after 15 days of culture in suspension. Cells within GelMA 5% hydrogels were collected after dissociation with collagenase type I. All samples were stored at −80 °C until RNA extraction. Total RNA was extracted using the Quick-RNA Microprep Kit (Zymo Research, Irvine, CA, USA) according to the manufacturer's protocol. RNA quantity and purity were assessed with a NanoDrop spectrophotometer (260/280 and 260/230 ratios), and RNA integrity was evaluated using an Agilent 2100 Bioanalyzer with an RNA 6000 Nano Kit. Only samples with RNA Integrity Number (RIN) greater than 7 were retained for library preparation RNA libraries were prepared using the AccuraCode RNA-Seq Kit (Cat. no. 10710174, Singleron Biotechnologies GmbH) following the manufacturer's instructions. Polyadenylated mRNA was isolated using AccuraCode beads (Cat. no. 1200030080). cDNA was synthesized with sample-specific barcodes and unique molecular identifiers (UMIs), pooled, and amplified to construct sequencing libraries.