Protocol to isolate and culture primary mouse feto-placental endothelial cells

Ionel Sandovici; Moritz Reiterer; Miguel Constância; Cristina M Branco

doi:10.1016/j.xpro.2022.101721

Protocol to isolate and culture primary mouse feto-placental endothelial cells

STAR Protoc. 2022 Dec 16;3(4):101721. doi: 10.1016/j.xpro.2022.101721. Epub 2022 Sep 24.

Authors

Ionel Sandovici¹, Moritz Reiterer², Miguel Constância³, Cristina M Branco⁴

Affiliations

¹ Department of Obstetrics and Gynaecology and National Institute for Health Research Cambridge Biomedical Research Centre, Cambridge CB2 0SW, UK; University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Institute of Metabolic Science, Addenbrookes Hospital, Cambridge CB2 0QQ, UK; Centre for Trophoblast Research, Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EG, UK. Electronic address: is299@cam.ac.uk.
² Physiological Laboratory, Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EG, UK; Patrick G Johnston Centre for Cancer Research, Queen's University Belfast, Belfast BT9 7AE, UK.
³ Department of Obstetrics and Gynaecology and National Institute for Health Research Cambridge Biomedical Research Centre, Cambridge CB2 0SW, UK; University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Institute of Metabolic Science, Addenbrookes Hospital, Cambridge CB2 0QQ, UK; Centre for Trophoblast Research, Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EG, UK.
⁴ Physiological Laboratory, Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EG, UK; Patrick G Johnston Centre for Cancer Research, Queen's University Belfast, Belfast BT9 7AE, UK. Electronic address: c.branco@qub.ac.uk.

Abstract

In the mouse, feto-placental endothelial cells (FPEC) line the inner surface of the feto-placental blood vessels located within placental labyrinthine zone and play critical roles in placental development and function. Here, we present a detailed protocol for isolation and culture of primary mouse FPEC, as well as two complementary methods (immunohistochemistry staining and flow cytometry analysis) to assess their purity. These cells are suitable for downstream ex vivo studies to investigate their functional properties, both in normal and pathological contexts. For complete details on the use and execution of this protocol, please refer to Sandovici et al. (2022).

Keywords: Cell biology; Cell culture; Cell isolation; Developmental biology; Flow cytometry/Mass cytometry; Microscopy; Tissue engineering.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Endothelial Cells*
Female
Flow Cytometry
Mice
Placenta*
Pregnancy

Abstract

Publication types

MeSH terms

Grants and funding