2025-12-14
ENA first public
ERC000011
ENA-CHECKLIST
Graduate Student, Department of Bioengineering, Indian Institute of Science
INSDC center name
public
INSDC status
ERS27355778
SRA accession
E-MTAB-16194:Sample 1
Submitter Id
8 to 12
week
age
ArrayExpress
broker name
White Blood Cells
cell type
not collected
collection date
house mouse
common name
Protocols: Mice were first anesthetized using high dose ketamine-xylazine solution. After anesthesia, about 500-1000 μL blood was collected through the retro-orbital vein. Following euthanasia, bronchoalveolar lavage fluid (BALF) was collected using a 20-G cannula was inserted into the trachea and fixed with a 93 ligature. Lungs were then lavaged with 1 mL sterile Ice-cold 1X phosphate buffered saline (PBS) two times (total volume 2 mL) to collect BALF. Fluid recovery was >85% each time. Blood and BALF samples were then centrifuged at 400 RCF for 5 mins at 4°C. The supernatant was stored at -80°C and the cell pellet was subjected to RBC lysis in ACK Lysis buffer and resuspended in 1mL PBS for counting using a hemacytometer. 1) Biomaterial implantation: Chitosan microspheres (337± 47 micrometers) were prepared using the Spraybase® Electrospray system inside a sterile biosafety cabinet and chemically sterilized as described22. Microspheres were surgically implanted into the peritoneal cavity of mice to induce chronic inflammation. In brief, microspheres (450 µL) were suspended in 450 µL sterile saline for implantation. The entire volume (900 µL) was then implanted into the peritoneal space22,27 following a laparotomy procedure. In controls (also called mock-surgery), the animals went through the surgical procedure and were injected with 900 µL sterile saline.  Immune cells from blood and BALF were isolated as described in earlier sections. Sample tagging was performed by labeling cells with BD® Mouse Single-Cell Multiplexing Kit. Neutrophils in the samples were tagged with Ly6G antibody-derived tags (Ly6G-ADTs, BD® AbSeq Ab-Oligos). These cells were subjected to single cell capture and cDNA Synthesis, library construction using a Whole Transcriptome Analysis (WTA, BD) kit with the BD Rhapsody™ single-cell analysis system following standard procedures provided by the manufacturer Sample tagging was performed by labeling cells with BD® Mouse Single-Cell Multiplexing Kit. Neutrophils in the samples were tagged with Ly6G antibody-derived tags (Ly6G-ADTs, BD® AbSeq Ab-Oligos). These cells were subjected to single cell capture and cDNA Synthesis, library construction using a Whole Transcriptome Analysis (WTA, BD) kit with the BD Rhapsody™ single-cell analysis system following standard procedures provided by the manufacturer.
description
mouse postnatal
developmental stage
healthy lung
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
Pool 1 (4 samples )
individual
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
Blood (Pooled)
organism part
Mus musculus
scientific_name
female
sex
Mock (ST01, ST02) or Chitosan Implanted in peritoneum (ST03,ST04)
stimulus
C57BL/6Ncrl
strain
Sample 1
title