# Protocol for isolating cervico-vaginal fluid cells from Macaca mulatta to study immunological and functional changes during pregnancy

## TL;DR

This paper provides a detailed protocol for isolating and analyzing cervico-vaginal fluid cells in rhesus macaques to study immune changes during pregnancy.

## Contribution

The paper introduces a novel, step-by-step method for studying immunological changes in cervico-vaginal fluid cells during pregnancy in Macaca mulatta.

## Key findings

- A protocol for cervico-vaginal fluid collection in pregnant rhesus macaques is described.
- Steps for cell isolation and immunophenotyping using flow cytometry are outlined.
- Protein-level detection in cervico-vaginal fluid is detailed using ELISA.

## Abstract

Cervico-vaginal fluid represents an important biological fluid due to its proximity to gestational tissues, such as the cervix, fetal membranes, and myometrium. Here, we present a protocol to isolate cervico-vaginal fluid cells from Macaca mulatta. We describe steps for cervico-vaginal fluid collection, cervico-vaginal cell isolation for nuclear staining and immunophenotyping by flow cytometry, and protein-level detection and quantification by ELISA. This protocol can provide insights into altered immunological changes at the cervico-vaginal interface.

•Instructions for optimal cervico-vaginal lavage collection in pregnant rhesus macaques•Protocol to characterize cervico-vaginal lavage cells by flow cytometry•Steps for protein-level detection in cervico-vaginal lavage by ELISA

Instructions for optimal cervico-vaginal lavage collection in pregnant rhesus macaques

Protocol to characterize cervico-vaginal lavage cells by flow cytometry

Steps for protein-level detection in cervico-vaginal lavage by ELISA

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Cervico-vaginal fluid represents an important biological fluid due to its proximity to gestational tissues, such as the cervix, fetal membranes, and myometrium. Here, we present a protocol to isolate cervico-vaginal fluid cells from Macaca mulatta. We describe steps for cervico-vaginal fluid collection, cervico-vaginal cell isolation for nuclear staining and immunophenotyping by flow cytometry, and protein-level detection and quantification by ELISA. This protocol can provide insights into altered immunological changes at the cervico-vaginal interface.