54 - Uterine Leukocyte Isolation and Culture

Abstract

Uterine leukocytes are a heterogeneous population that increase dramatically in number during pregnancy. A transient accumulation of leukocytes occurs in the decidua basalis of the maternal–fetal interface. The early primary leukocyte populations are the subsets of uterine natural killer (uNK) cells, whereas dendritic cells, macrophages, and T lymphocytes represent minor populations. The functions of the cells of each of these lineages in the pregnant uterus are intriguing questions for reproductive immunologists. Unlike studies of cells from many other organs, in vitro studies of cells isolated from the pregnant mouse uterus as freshly isolated leukocytes or longer-term cultured cells are still performed poorly. This is because there is no established protocol that has been generally useful in such experiments. Indeed, the dynamic changes and complexity of tissues from the gestational uterus limits a conventional approach and requires an advanced protocol to achieve reproducible results. Strong culture protocols or uterine immune cell lines would represent a major advance for studies of the immunoregulatory events occurring during pregnancy. In this chapter, we discuss critical points that must be considered during uterine tissue sampling due to the stage of pregnancy at the time of uterine leukocyte isolation. We also outline our method for selective purification and culture of leukocyte subsets, using uNK cells as an example.