细胞技术

Over the past two decades there have been unprecedented advances in the capabilities for live cell imaging using light and confocal microscopy. Together with the discovery of green fluorescent protein and its derivatives and the development of a vast array of fluorescent imaging probes a ...

In order to understand osteoclast cell biology, it is necessary to culture these cells on a physiological �substrate that they can resorb in vitro, such as bone or dentine. However, this creates problems for analysis by fluorescence microscopy, due to the depth of the sample under investigation. ...

This chapter describes the use of digital micro-radiography with the Faxitron machine as a means of imaging and quantitating bone mineral content in mice and rats.

This chapter provides information for the use of peripheral quantitative computed tomography in small animals, including suggestions for study design, instrument setting, and data interpretation.

This chapter describes the use of microcomputed tomography scanning for analysing bone structure, focussing on rodent bone. It also discusses sample preparation, the correct set-up of the scanner, and the impact of some of the important scanner settings.

Optical imaging is a valuable technique for visualizing and quantifying biological processes in living �organisms. Optical imaging can be divided into two main imaging modalities: bioluminescence imaging and fluorescence imaging. This chapter describes the use of these imagi ...

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 ...