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A simple and reliable method to measure proteoglycan synthesis by chondrocytes in culture is described. Confluent chondrocytes in 24-well plates are labeled for 24–72 h with 35SO4 2− in the presence of stimulating agents. At the end of treatment, the secretion medium containing radiolabel ...

Cultures of cartilage explants have long been used to study the effects of modulators of extracellular matrix degradation. We present a simple and rapid assay system, based on culture of rabbit cartilage explants, which permits study of the effects of protease inhibitors on proteoglycan d ...

The use of synthetic peptides to generate rabbit polyclonal anticatabolic neoepitope antibodies that can be used to study the presence of defined proteolytic cleavage sites in aggrecan is described. Principles of peptide design and methods for preparation and characterization of ...

Apoptosis is a form of programmed cell death conceptually opposed to necrosis. In view of the inherent difficulty in accurately detecting apoptosis in chondrocytes, this chapter describes complementary techniques that may be used in combination. During apoptotic death, protein and ...

Quantitative immunoassays have been developed to measure the content, degradation, and synthesis of types II and IX collagens in hyaline cartilages. Some of these assays and their applications are described in this chapter. These and other assays are commercially available. The appli ...

The placenta has multiple functions that are vital to the success of pregnancy. Trophoblast cells are specialized constituents of the placenta that are responsible in large part for these unique functions. In some species, trophoblast cells acquire invasive properties. These cells pe ...

In vivo analysis of trophoblast cell invasion is highly dependent on histological techniques, which are amply described in standard textbooks. The emphasis of this chapter therefore lies on material collection and interpretation of tissue sections, rather than on histological te ...

Extravillous trophoblasts invade the uterine wall (interstitial invasion) and the spiral arteries (endovascular invasion), replacing the cells of the vessel wall and creating a high-flow, low-resistance vessel. We describe a model to allow the interactions between the invading t ...

Two methods are described for the study of human trophoblast invasion. When first-trimester placental villi are explanted on gels of a permissive extracellular matrix (ECM), a population of pure extravillous trophoblast cells grows out during the following several days from villous t ...

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 ...