By Michael J. Soares, Joan S. Hunt
A set of state of the art laboratory concepts for the learn of the trophoblast and placental biology. The ideas provided diversity from experimental animal types, to animal and human placental organ and phone tradition structures, to morphological, biochemical, and molecular options for assessing trophoblast/placental development, differentiation and serve as. quantity 2 offers without difficulty reproducible protocols for learning trophoblast invasion, maternal immune telephone responses to being pregnant, and the hormone generating services of the placenta. There also are the right way to examine the barrier functionality of the placenta, its function in regulating the delivery of meals and wastes, and the diversifications of trophoblast cells and the placenta to illness. Highlights comprise suggestions to investigate trophoblast mobilephone interactions with the uterine vasculature and immune cells, and the endocrine capabilities of the placenta. A spouse first quantity concentrates on equipment for examine embryo-uterine implantation, trophoblast phone improvement, and the association and molecular characterization of the placenta.
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Additional info for Placenta and Trophoblast Volume 2 Methods And Protocols (Methods in Molecular Medicine)
1. 1. FIXATION, PARAFFIN EMBEDDING, AND SECTIONING Hysterectomy material offers technical and logistic problems for processing and sectioning, and proper fixation is a particular problem. As a first and necessary step, the amniotic fluid should be removed using a syringe and replaced by fixative. Large organs such as a pregnant uterus must remain immersed in fixative for a sufficiently long time in order to obtain satisfactory fixation. The use of an easily penetrating fixative such as buffered (para)formaldehyde is therefore necessary.
STAINING For many years, placental bed work was bedeviled by the virtual impossibility to distinguish with certainty the invading trophoblast from decidual cells, unless, of course, one wished to concentrate on the multinuclear giant cells. That was also the reason why Pijnenborg and colleagues (9) had restricted their quantitative work on interstitial trophoblast to the myometrium, where it was assumed that the trophoblast cells could reasonably well be distinguished from the spindle-shaped smooth muscle cells.
Many diseases of pregnancy, including preeclampsia, impact nutrient flow to the placenta and fetus. One of the essential nutrients is oxygen. Chapter 27 presents in vitro methods for studying the effects of oxygen on trophoblast cell development. This chapter describes protocols for studying trophoblast cell adaptive responses to restricted oxygen and following exposure to oxidative stress. In Chapter 28, a simple procedure is outlined for exposing pregnant rodents to hypoxia. Either rats or mice are easily placed into a hypobaric chamber, which simulates high altitude and creates a hypoxic atmosphere.
Placenta and Trophoblast Volume 2 Methods And Protocols (Methods in Molecular Medicine) by Michael J. Soares, Joan S. Hunt