TEB
Mus musculus
Protocols: Experimental animals were obtained by breeding of the parental strains, the genotypes were determined by genotyping. The mice were housed in individually ventilated or open cages, all with ambient temperature of 22°C, a 12 h:12 h light:dark cycle, and food and water ad libitum. Microdissected pieces of mammary gland pooled from 8 animals were digested in a collagenase/hyaluronidase solution [3 mg/ml collagenase A, 100 U/ml hyaluronidase (all Merck), 5% fetal bovine serum (FBS; Hyclone/GE Healthcare) in CO2 independent medium (Thermo Fisher Scientific)] for 2 h at 37°C, shaking at 120 rpm. Resulting tissue fragments were treated with 0.25% trypsin, 5 mg/ml dispase II, 100 µg/ml DNase and red blood cell lysis buffer (Sigma) and filtered through a 40 µm cell strainer. Resulting single cell suspension was spun down and resuspended in flow buffer (5 mM EDTA, 1% BSA, 1% FBS in phenol red-free DMEM/F12 with HEPES) for single cell RNA sequencing and filtered through a 40 µm cell strainer. The analysis was done on Fusion (BD), operated by SORP software (BD). The resulting cell pellet was pooled down, resuspended in freezing medium (10% DMSO in FBS) and frozen by slow freezing at -80°C and moved to liquid nitrogen 24 h thereafter. For microdissection of TEB- or duct-containing regions, 5 weeks old R26-mTmG females were used. Mammary glands #3 and #4 were dissected and spread on 10 cm plastic petri dishes. Under the control of a fluorescent stereoscope (Leica FM165C), 0.5×0.5 mm pieces of tissue containing either TEBs or pieces of ducts (assessed based on tdTomato fluorescence) were dissected and collected in CO2 independent medium on ice. 5-8 mice were pooled for one experiment.  Single-cell suspensions were prepared and processed using the GEM-X Single Cell 3' v4 kit (10x Genomics) following the manufacturer's protocol for frozen cells. Nucleic acid libraries were prepared and processed using the GEM-X Single Cell 3' v4 kit (10x Genomics) following the manufacturer's protocol for frozen cells.
ENA first public
2025-08-07
INSDC center name
Institute of Molecular Genetics of the Czech Academy of Sciences
INSDC status
public
Submitter Id
E-MTAB-15259:TEB
age
5
broker name
ArrayExpress
cell type
all cells
collection date
not collected
common name
house mouse
developmental stage
puberty
genotype
R26-mTmG homozygous
geographic location (country and/or sea)
not collected
individual
NA
isolate
not applicable
organism part
mammary gland TEB region
sample name
E-MTAB-15259:TEB
scientific_name
Mus musculus
sex
female
strain
R26-mTmG