GSM8988041: Day 14 Healthy Control, Spleen, scRNAseq; Mus musculus; RNA-Seq
Female Balb/c mice (7–8 weeks old) were orthotopically inoculated with 2,000,000 4T1-tdTomato-luc2 cells into the fourth right mammary fat pad. Healthy control mice did not receive tumor cell inoculation. At 7, 14, or 21 days post-inoculation, both tumor-bearing and control mice were euthanized via CO₂, and their lungs, spleens, and biomaterial scaffolds were surgically explanted for analysis. Single-cell suspensions prepared from the isolated tissues were processed using a Drop-Seq device (FlowJEM; Toronto, CA), following the protocol described by Macosko et al., 2015. The device was run for 10 minutes, after which the resulting cell-bead pairs (STAMPs) were collected and the oil phase was removed. Approximately 1,000 STAMPs were subjected to reverse transcription, Exonuclease I treatment, and universal PCR amplification. cDNA concentration and quality were assessed using a Bioanalyzer with a DNA High Sensitivity chip (Agilent Technologies; Santa Clara, CA). cDNA tagmentation was performed using Tn5 transposase, as described by Picelli et al., 2014.