细胞功能测定

Raman microscopy is a non-destructive technique requiring minimal sample preparation that can be used to measure the chemical properties of the mineral and collagen parts of bone simultaneously. Modern Raman instruments contain the necessary components and software to acquire the ...

Fourier transform infrared imaging (FTIRI) is a technique that can be used to analyze the material properties of bone using tissue sections. In this chapter I describe the basic principles of FTIR and the methods for capturing and analyzing FTIR images in bone sections.

This chapter describes the surgical procedures for ovariectomy and orchidectomy in mice and rats. In �addition to providing technical details of the surgical techniques, details of anaesthesia and perioperative care are also included.

This chapter describes the calvarial injection method, whereby the effect of a substance on bone is tested by subcutaneous injection over the calvarium of a mouse. This assay allows testing of the effect of substances on both bone resorption and bone formation in a relatively simple in vivo model. ...

The primary functions of bone are to do with support and protection – mechanical functions. The aim of this chapter is to set out some of the methods that can be used to measure these properties in cortical and cancelleous bone from large (e.g. human or bovine) and small (e.g. mouse) animals. The difference bet ...

Bone cells of the osteoblastic lineage are responsive to the local mechanical environment. Through integration of a number of possible loading-induced regulatory stimuli, osteocyte, osteoblast, and osteoclast behaviour is organized to fashion a skeletal element of sufficient ...

This chapter describes several methods suitable for mechanically stimulating monolayers of bone cells by fluid shear stress (FSS) in vitro. Fluid flow is generated by pumping culture medium through two parallel plates, one of which contains a monolayer of cells. Methods for measuring nit ...

The skeleton fulfils its mechanical functions through structural organisation and material properties of individual bones. Both cortical and trabecular morphology and mass can be (re)modelled in response to changes in mechanical strains engendered by load-bearing. To address ...

This chapter describes the isolation, culture and staining of primary osteoblasts from the calvaria and long bones of neonatal rats. The key advantages of this assay are that it allows direct measurement of bone matrix deposition and mineralisation, as well as yielding good quantities of os ...

This chapter describes the isolation of primary mouse osteoblasts from adult mouse calvaria and long bones, as well as the process of isolation of bone cells from neonatal mouse calvaria. Osteoblasts from adult mouse bone are obtained as outgrowth from collagenase-treated bone pieces. Is ...

Osteoblast cultures can be used to investigate the mechanisms of bone formation, to probe the cellular and molecular basis of bone disease, and to screen for potential therapeutic agents that affect bone formation. Here, we describe the methods for establishing and characterising prima ...

Osteocytes are the terminally differentiated cells of the osteoblastic lineage embedded within the mineralized bone matrix. They have been identified as key players in mechanotransduction and in mineral and phosphate homeostasis. In addition, they appear to have a role in mediating ...

Osteocytes can be isolated from chicken calvaria using mild EDTA treatment alternating with collagenase treatment. The cell population obtained contains both osteoblasts and osteocytes. A pure population of osteocytes is obtained following immunomagnetic separation with ...

We describe the culture and use of MLO-Y4 cells in studies of gene expression, response to fluid flow, and dendrite growth. We also describe how to use the MLO-A5 cells as a model of osteoblast to osteocyte �differentiation and how to study their mineralization. These studies serve as a beginning point to ...

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 ...