2025-08-01
ENA first public
ERC000011
ENA-CHECKLIST
Department of Internal Medicine, Division of Gastroenterology and Hepatology, University of Michigan Medical School
INSDC center name
public
INSDC status
ERS23860290
SRA accession
E-MTAB-14975:HT-561-Distal
Submitter Id
120
day
age
ArrayExpress
broker name
not collected
collection date
human
common name
Protocols: Human lung tissue research was reviewed and approved by The University of Michigan Institutional Review Board (IRB). Normal, de-identified human fetal lung tissue was obtained from the University of Washington Laboratory of Developmental Biology. Tissue was shipped overnight in UW-Belzer’s solution (Thermo Fisher, Cat#NC0952695) on ice and was processed for experiments or fixation within 24h. Normal, de-identified, donor human adult lungs not eligible for transplant were obtained from Gift of Life Michigan Donor Care Center. Specific tissue regions were dissected according to anatomic region and snap-frozen in liquid nitrogen for further downstream analysis. Preparation was performed as described previously in accordance with 10X Genomics’ protocols (CG000375 Rev B for fetal tissue). Briefly, snap-frozen tissue was minced into smaller fragments using a scalpel and then added to lysis buffer (Thermo Fisher Scientific, Cat#PI28324). Tissue was homogenized using a pellet pestle 15 times then incubated for 5 minutes in lysis buffer. The cell suspension was filtered through a 33 μm strainer. Suspension was centrifuged at 500g for 5 mins at 4°C. Supernatant was removed and the pellet was washed with PBS + 1% BSA, centrifuged at 500g for 5 mins at 4°C; pellet was washed and centrifuged again. An added debris removal column step was used for the adult tissue.  Permeabilization for ATAC-seq samples was performed by incubating the pellet in 0.1X lysis buffer and incubated for 2 minutes. The suspension was then centrifuged at 500g for 5 mins at 4°C; supernatant was removed and pellet was resuspended in diluted nuclei buffer. Immediately after tissue dissociation, single nucleus libraries were prepared on the Chromium Next GEM Single Cell Multiome ATAC + Gene Expression Reagent Bundle, 16 rxns PN-1000283 was used with a target of 7500 cells.
description
fetus
developmental stage
not collected
geographic location (country and/or sea)
not applicable
isolate
Late pseudoglandular
lung developmental stage
Homo sapiens
organism
Homo sapiens
organism
lung
organism part
Distal
position
lung, distal
sampling site
Homo sapiens
scientific_name
female
sex
HT-561-Distal
title