An Efficient Method for Extracting Human Fallopian Tube Epithelia for Single-cell Analyses

Human fallopian tubes are intrinsic to reproduction. The purpose of the fallopian tubes is to allow for the transit of sperm, the ovum, and if fertilization is successful, the embryo. The epithelial cells that line the inner surface of fallopian tubes are integral to both normal and abnormal fallopian tube processes, including disease initiation. After menopause, fallopian tubes cease to have a significant role in the body and the intraepithelial cell makeup changes. We describe a method in which these epithelial cells can be isolated from fresh fallopian tubes with minimal stromal cell contamination into a single-cell suspension. These cells can be grown in culture or used for further analysis such as flow cytometry and single-cell RNA sequencing. This isolation protocol can be achieved in 4-6 h and yields viable cells that can be used for immediate downstream analysis. This efficient protocol facilitates the isolation of fallopian tube epithelial cells with an enriched epithelial population.

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Publication Details

Journal
Journal of Visualized Experiments
Published
2025-03-28
DOI
https://doi.org/10.3791/67753
Primary Topic
Single-cell and spatial transcriptomics
Type
article
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article

An Efficient Method for Extracting Human Fallopian Tube Epithelia for Single-cell Analyses

Laura B. James‐Allan, Lili Yang, Sanaz Memarzadeh, Gabriella DiBernardo et al.
1 links
Journal of Visualized Experiments
Single-cell and spatial transcriptomics
article

An Efficient Method for Extracting Human Fallopian Tube Epithelia for Single-cell Analyses

Laura B. James‐Allan, Lili Yang, Sanaz Memarzadeh, Gabriella DiBernardo, Christopher J Ochoa, Yan-Ruide Li, Lauryn Ruegg
article en

Abstract

Human fallopian tubes are intrinsic to reproduction. The purpose of the fallopian tubes is to allow for the transit of sperm, the ovum, and if fertilization is successful, the embryo. The epithelial cells that line the inner surface of fallopian tubes are integral to both normal and abnormal fallopian tube processes, including disease initiation. After menopause, fallopian tubes cease to have a significant role in the body and the intraepithelial cell makeup changes. We describe a method in which these epithelial cells can be isolated from fresh fallopian tubes with minimal stromal cell contamination into a single-cell suspension. These cells can be grown in culture or used for further analysis such as flow cytometry and single-cell RNA sequencing. This isolation protocol can be achieved in 4-6 h and yields viable cells that can be used for immediate downstream analysis. This efficient protocol facilitates the isolation of fallopian tube epithelial cells with an enriched epithelial population.

Journal of Visualized Experiments(217)
University of California, Los Angeles (US), Parker Institute for Cancer Immunotherapy (US), Broad Center (US), VA Greater Los Angeles Healthcare System (US)
Zero hunger
Openalex Percentile: Top 95%
Single-cell and spatial transcriptomics
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