Protocol for isolating amnion cells from human and non-human primate placenta for flow cytometry and transcriptomics

Monica Cappelletti; Daniel Short; Marco Morselli; Matteo Pellegrini; Mark R Johnson; Yalda Afshar; Suhas G Kallapur; Pietro Presicce

doi:10.1016/j.xpro.2024.103044

Protocol for isolating amnion cells from human and non-human primate placenta for flow cytometry and transcriptomics

STAR Protoc. 2024 Apr 27;5(2):103044. doi: 10.1016/j.xpro.2024.103044. Online ahead of print.

Authors

Monica Cappelletti¹, Daniel Short², Marco Morselli³, Matteo Pellegrini⁴, Mark R Johnson², Yalda Afshar⁵, Suhas G Kallapur¹, Pietro Presicce⁶

Affiliations

¹ Divisions of Neonatology and Developmental Biology, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA.
² Department of Metabolism, Digestion and Reproduction, Imperial College London, London, UK.
³ Department of Chemistry, Life Sciences and Environmental Sustainability (S.C.V.S.A.), University of Parma, 43124 Parma, Italy.
⁴ Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA 90095, USA.
⁵ Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA 90095, USA; Division of Maternal Fetal Medicine, Department of Obstetrics and Gynecology, David Geffen School of Medicine at the University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁶ Divisions of Neonatology and Developmental Biology, David Geffen School of Medicine at the University of California Los Angeles, Los Angeles, CA 90095, USA. Electronic address: ppresicce@mednet.ucla.edu.

Abstract

The amnion is a thin layer of fetal origin in contact with the amniotic fluid which plays a key role at the feto-maternal interface during pregnancy. Here, we present a protocol for isolation of human and Rhesusmacaque amnion cells. We describe steps for tissue dissection, cell isolation for flow cytometry analysis, and RNA isolation for RNA sequencing library preparation and analysis. This protocol can provide insights into altered immunological pathways during intrauterine infections to develop new therapeutic strategies. For complete details on the use and execution of this protocol, please refer to Presicce et al.¹.

Keywords: Flow Cytometry; Immunology; Model Organisms; Sequence analysis.