A wide variety of techniques has been developed for qualitative and quantitative analysis of gene expression in human cells and tissues. Two commonly used methods are reverse-transcription (RT)-polymerase chain reaction (PCR) to analyze the transcribed messenger RNAs (mRNA) and im ...

The human major histocompatibility complex (MHC) contains genes encoding the Human Leukocyte Antigens (HLA). Of these antigens, placental immunologists need study only the HLA class I molecules, because HLA class II expression is repressed in the fetal placental cells that are in direct ...

Decidualization of the mouse and human uterus is accompanied by the influx of large numbers of natural killer lymphocytes (uterine natural killer cells). Adoptive cell transfer to mated, alymphoid mice is a general model suitable for analysis of homing, differentiation and function of the ...

To promote the use of the nonhuman primate model for the study of the cellular and molecular biology of maternal-fetal interactions and placental development during early pregnancy, we have developed protocols for the isolation and characterization of placental trophoblasts and de ...

Human monocytes and macrophages, which are also called mononuclear phagocytes, represent a major arm of the innate immune system. These cells not only protect against infection but are also central to tissue remodeling and production of chemokines, cytokines, and growth factors. Tissue ...

During human pregnancy, fetal placental cells known as trophoblast invade into the uterine mucosal lining, coming into contact with maternal cells including a specialized population of leukocytes. In order to understand the interaction of maternal cells with trophoblast, it is usef ...

During implantation and pregnancy, the invading trophoblast population is within close contact to maternal immune cells, particularly macrophages. During this period, a low level of trophoblast cell death occurs as part of the normal process of tissue renewal. Macrophage engulfme ...

This chapter presents experimental methods to measure in vivo uteroplacental nutrient substrate uptake, transfer to the fetus, and metabolism by the uteroplacental unit. The fundamental method involves application of the Fick principle, which requires determination of uteri ...

In vitro models have proven to be effective in studying the placental transporters that play a role in the exchange of nutrients, waste products, and drugs between the maternal and fetal circulations. Although primary cultures of trophoblast cells can be used to perform uptake, efflux, and met ...

Isolated plasma membrane vesicles from human placenta allow transporter-mediated mechanisms across individual plasma membranes to be identified and characterized in vitro. This approach is reliant on isolating each of the trophoblast plasma membranes, either the maternal ...

A method is described for measuring amino acid uptake by the syncytiotrophoblast of the human placenta in vitro using taurine as an example. Small fragments of placental villous tissue (2–3 mm3) are tied to a comb that enables them to be moved in concert between a series of incubation and wash buffers. The ...

Fatty acids (FAs), especially essential fatty acids (EFAs) and their long chain polyunsaturated fatty acid (LCPUFAs) derivatives, are critical for proper fetal development. The fetus relies on the placental transfer of EFAs from the maternal circulation for development. In fact, fatty ...

We use two different heterologous expression systems to characterize the functional features of plasma membrane transporters cloned from placenta. The first is the vaccinia virus expression system that utilizes a recombinant vaccinia virus carrying a transgene for T7 RNA polyme ...

Placental progesterone synthesis in humans prevents abortion of the fetus by maintaining uterine quiescence and low myometrial excitability. In rodents, a transient steroidogenic output is observed in the trophoblast giant cells during mid-pregnancy. Although the exact role of ...

The pregnancy-associated glycoproteins (PAGs) are a large gene family expressed in trophoblast cells of ruminant ungulates. The detection of PAGs in maternal serum has served as the basis for pregnancy detection in ruminant ungulates and also for use as markers of trophoblast developm ...

In this chapter, we describe protocols for the generation and characterization of alkaline phosphatase-ligand fusion proteins and their use as tools for the identification of specific ligand-receptor interactions.

Complementary DNAs of three recombinant proteins related to the prolactin family: ovine placental lactogen (oPL), ovine prolactin (oPRL), and rabbit soluble extracellular domain of prolactin receptor (rbPRLR-ECD) were subcloned by different methods and inserted into prokar ...

Placental hormones contribute to changes in maternal physiology, especially to changes in the blood system. Methods are described to express a placental hormone from a cloned cDNA by transfection into a mammalian cell line, to purify the hormone, and to assess the activities of the hormone in pr ...

Interferon (IFN)τ, a Type I IFN produced by the conceptus trophectoderm, is the pregnancy recognition signal in ruminants that prevents development of the endometrial luteolytic mechanism. In addition, IFNτ acts in a paracrine manner on the ovine endometrium to induce or increase expre ...

Despite being one of the leading causes of maternal death and a major contributor of maternal and perinatal morbidity, the mechanisms responsible for the pathogenesis of preeclampsia are unknown. The initiating event in preeclampsia has been postulated to be reduced uteroplacental p ...