Purification of primary human placental leukocytes to study maternal-fetal interactions

Nadia M Ikumi; Zachary Koenig; Shweta Mahajan; Clive M Gray; Tamara Tilburgs

doi:10.1016/j.xpro.2023.102277

Purification of primary human placental leukocytes to study maternal-fetal interactions

STAR Protoc. 2023 May 12;4(2):102277. doi: 10.1016/j.xpro.2023.102277. Online ahead of print.

Authors

Nadia M Ikumi¹, Zachary Koenig², Shweta Mahajan², Clive M Gray³, Tamara Tilburgs⁴

Affiliations

¹ Division of Immunology, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Department of Obstetrics and Gynaecology, University of Cape Town, Cape Town, South Africa. Electronic address: nadia.ikumi@uct.ac.za.
² Division of Immunobiology, Center for Inflammation and Tolerance, Cincinnati Children's Hospital, Cincinnati, OH 45229, USA; Immunology Graduate Program, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA.
³ Division of Immunology, Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Division of Molecular Biology and Human Genetics, Stellenbosch University, Cape Town, South Africa.
⁴ Division of Immunobiology, Center for Inflammation and Tolerance, Cincinnati Children's Hospital, Cincinnati, OH 45229, USA; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA. Electronic address: tamara.tilburgs@cchmc.org.

Abstract

Decidual leukocytes play key roles in maternal-fetal tolerance and immunity. Here, we present detailed methods to purify, culture, and functionally analyze human placental dNK, dTreg, dTem, and dMɸ from decidua parietalis, the maternal part of the placental membranes; decidua basalis, the maternal part of the placenta; and placental villi. These sites have high clinical relevance in the development of villitis and chorioamnionitis. This allows in-depth phenotypic and functional investigation of placental immune populations and their interactions with extravillous trophoblasts. For complete details on the use and execution of this protocol, please refer to Ikumi et al.,¹ Tilburgs et al.,² Salvany-Celades et al.,³ Crespo et al.,⁴ van der Zwan et al.⁵.

Keywords: Cell Biology; Cell culture; Cell isolation; Cell separation/fractionation; Developmental biology; Immunology.