Evidence map›Paper›PMID 37602769›Full record

ArticleThe Laryngoscope2024

Method for Collecting Single Epithelial Cells from the Mouse Larynx.

Mohammed Imran Khan, Meena Easwaran, Joshua D Martinez, Akari Kimura, Elizabeth Erickson-DiRenzo

Open access · bronzeAbstract read
In one paragraph

Article in The Laryngoscope, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
0.3field-weighted citation impact, top 36% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed, 1 citations in OpenAlex.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors at 1 institution in 1 country.

Mohammed Imran KhanDepartment of Otolaryngology - Head & Neck Surgery, School of Medicine, Stanford University, Stanford, California, U.S.A.ORCID 0000-0003-3020-1324
Meena EaswaranDepartment of Otolaryngology - Head & Neck Surgery, School of Medicine, Stanford University, Stanford, California, U.S.A.
Joshua D MartinezDepartment of Otolaryngology - Head & Neck Surgery, School of Medicine, Stanford University, Stanford, California, U.S.A.
Akari KimuraDepartment of Otolaryngology - Head & Neck Surgery, School of Medicine, Stanford University, Stanford, California, U.S.A.ORCID 0000-0001-6682-8212
Elizabeth Erickson-DiRenzoDepartment of Otolaryngology - Head & Neck Surgery, School of Medicine, Stanford University, Stanford, California, U.S.A.ORCID 0000-0002-9841-0184
Stanford Medicine · US

Funding

Evaluating the role of epithelial basal cells in laryngeal homeostasis and disease developmentR01DC020176 · NIDCD · STANFORD UNIVERSITY · PI Elizabeth DiRenzo · 2022 to 2026
$1.9M
Ultra high throughput sequencer for sustaining and enhancing multi-scale genomic studiesS10OD025212 · OD · STANFORD UNIVERSITY · PI SNYDER, MICHAEL P. · 2018 to 2018
$600k
Biological and clinical evaluation of the laryngeal mucus layerR21DC016126 · NIDCD · STANFORD UNIVERSITY · PI DIRENZO, ELIZABETH · 2018 to 2020
$565k
Computer ClusterS10OD021763 · OD · STANFORD UNIVERSITY · PI JI, HANLEE P · 2016 to 2016
$481k
NIDCD NIH HHS R01 DC020176NIDCD NIH HHS R21 DC016126NIH HHS 1S10OD021763NIH HHS S10 OD021763NIH HHS S10 OD025212NIH HHS S10OD025212
6 · The paper itself

Abstract

objectiveThe larynx is lined by specialized epithelial cell populations. Studying molecular changes occurring in individual epithelial cell types requires a reliable method for removing these cells from the larynx. Our objective was to develop a method to harvest individual epithelial cells from the mouse larynx while minimizing contamination from non-laryngeal sites and non-epithelial laryngeal cells.

methodsMice were euthanized, and the larynx was carefully exposed and separated from non-laryngeal sites. A small dental brush was inserted into the laryngeal inlet and rotated to obtain epithelial cells. Cells were transferred to collection media, counted, and cytospin preparations stained for laryngeal epithelial (i.e., Pan-Keratin, EpCAM, NGFR, p63, K5, β-tubulin, MUC5AC) and non-epithelial (i.e., vimentin) cell markers. Histopathology was completed on brushed laryngeal tissue sections to evaluate the depth of cell collection. Preliminary Single-cell RNA sequencing (scRNA-seq) was performed to confirm this method can capture diverse laryngeal cell types.

resultsWe collected 6000-8000 cells from a single larynx and 35000-40000 cells from combining brushings from three tissues. Histopathology demonstrated brushing removed the epithelial layer of the larynx and some underlying tissue. Immunofluorescence staining demonstrated the phenotype of harvested cells was primarily epithelial. Preliminary scRNA-seq was successfully conducted and displayed nine unique cell clusters.

conclusionWe developed a reliable method of harvesting individual epithelial cells from the mouse larynx. This method will be useful for collection of laryngeal cells for a variety of downstream cellular and molecular assays, including scRNA-seq, protein analyses, and cell-culture-based experiments, following laryngeal injury. LEVEL OF EVIDENCE: NA Laryngoscope, 134:786-794, 2024.

Indexed as

LarynxAnimalsCell Culture TechniquesEpithelial CellsMiceimmunofluorescencelarynxmouse larynx epithelial cellsscRNA-seq

Identifiers

PMID37602769
PMCPMC10841475
OpenAlexW4386019827

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.