细胞技术

In this chapter, we describe techniques for the isolation and characterization of skeletal stem cells from human bone marrow. The methods for enrichment of STRO-1 positive cells using magnetic activated cell sorting are described and we also cover techniques for establishing and chara ...

Newborn rabbits provide a useful and readily available source of authentic mature osteoclasts, which can be easily isolated directly from the long bones in relatively large numbers, compared to other rodents. Primary cultures of authentic rabbit osteoclasts on resorbable substra ...

The murine co-culture assay is used to generate mature osteoclasts from bone marrow precursors by culturing them with osteoblasts that are stimulated with 1,25-dihydroxy vitamin D3 and prostaglandin E2. This assay is used particularly to analyse osteoblast–osteoclast interact ...

Osteoclasts are multi-nucleated cells that have the unique ability to resorb calcified bone matrix. They derive from haematopoietic precursor cells, and can be generated in vitro by stimulation of peripheral blood mononuclear cells with the cytokines M-CSF and RANKL. In this chapter, we d ...

Extensive research efforts over the years have provided us with great insights into how bone-resorbing osteoclasts (OCs) develop and function and, based on such work, valuable antiresorptive therapies have been developed to help combat the excessive bone loss that occurs in numerous sk ...

This chapter describes quantitative methods for isolating and culturing rodent osteoclasts on dentine, a bone-like, resorbable substrate. These techniques generate relatively large numbers of osteoclasts and allow the key processes of osteoclast formation and activation to ...

Osteoclasts originate from hematopoietic myeloid progenitors that differentiate into specialized multinucleated cells uniquely capable of resorbing bone in both physiological and pathological conditions. Osteoclast numbers and degradative activities increa ...

The important roles of lipids particularly certain phospholipids in signal transduction processes and as important disease markers are becoming increasingly evident. Unfortunately, however, sensitive methods of lipid analysis are established to a much lesser extent than, e.g ...

The use of NMR spectroscopy in lipid research has been traditionally reserved for the analysis and structural elucidation of discrete lipid molecules. Although NMR analysis of organic molecules provides a plethora of structural information that is normally unattainable by most ot ...

A gradient HPLC-charged aerosol detection method was applied to the measurement of different lipids including: free fatty acids, fatty alcohols, glycerides, steroids, phospholipids, and fat-soluble vitamins. An algal oil sample is used as an example. Charged aerosol detection offe ...

There has been a recent explosion in research concerning novel bioactive sphingolipids (SPLs) such as ceramide (Cer), sphingosine (Sph), and sphingosine 1-phosphate (Sph-1P) and this has necessitated the development of accurate and user-friendly methodology for analyzing and qu ...

Modern lipidomics relies heavily on mass spectrometry for the structural characterization and quantification of lipids of biological origins. Structural information is gained by tandem mass spectrometry (MS/MS) whereby lipid ions are fragmented to elucidate lipid class, fat ...

Every organ compromises of several different cell types. When studying the effects of a chosen compound within this organ or tissue uptake, localisation, metabolism, and the effect itself can be expected to differ between cells. Using the example of Vitamin E in pulmonary tissue we introduce c ...

Lipases are responsible for the hydrolysis of acylglycerols and cholesteryl esters in animals, plants, and microorganisms. In this chapter we describe a tool for the concomitant analysis of lipases in complex proteomes. For this purpose, the target enzymes are selectively and covalen ...

Plant oil has become an important component in the search for a replacement for non-renewable energy sources, as well as being used for a wide range of industrial purposes, all in addition to its vital importance for human diet. Detailed knowledge of the lipid distribution in plants is fundamental f ...

In the last decades, free radical processes delineated an interdisciplinary field linking chemistry to biology and medicine. Free radical mechanisms became of importance as molecular basis of physiological and pathological conditions. Lipids, in particular, unsaturated fa ...

Very long chain fatty acids confer functional diversity on cells by variations in their chain length and degree of unsaturation. Microsomal fatty acid elongation represents the major pathway for determining the chain length of saturated, monounsaturated, and polyunsaturated fa ...

Quantitative analysis of components from different subcellular fractions is a key to the understanding of metabolic function as well as to the origin, the biogenesis, and the crosstalk of organelles. The yeast is an excellent model organism to address such questions from the biochemical, m ...

Adipose tissue development undergoes remodeling in terms of newly synthesized cells (hyperplasia) and newly synthesized lipids that accumulate in adipocytes (hypertrophy). Synthesis and/or breakdown rates of adipose cells and lipids follow a continuous and dynamic pattern, e ...

Vegetable oils are complex mixtures containing a wide range of major compounds. Triacylglycerols (TAGs; consisting of a glycerol moiety with each hydroxyl group esterified to a fatty acid) are the major components (95–98%) of vegetable oils. TAGs are an important source of energy and nutrit ...