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Title: Isolation, generation and characterization of equine osteoclasts
Author: Gray, A.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2001
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a) Equine osteoclasts (OCs) were identified in ex vivo samples of developing long bone. They were particularly numerous in areas of active bone formation and remodelling such as at the longitudinal septum between cartilage and subchondral bone and in the spongiosum. Equine OCs were identified as being typically multinuclear and expressing very high levels of tartrate resistant acid phosphatase (TRAP) and cathepsin K. Only low levels of cathepsin B were detected in equine OCs. b) Techniques were developed to isolate equine OCs for the first time in vitro. The phenotype of these cells was clearly defined; they were multinuclear, expressed TRAP, the vitronectin receptor (VNR) and were observed in direct association with areas of resorption when cultured on ivory slices. However, equine OCs could only be isolated in small numbers in vitro and were often of poor viability. c) Techniques were developed to generate equine osteoclast-like cells (OCLs) for the first time in vitro from bone marrow cells (BMCs) with or without osteoblastic support cells. d) Equine OCLs demonstrated high levels of cathepsin K activity but only low levels of cathepsin B activity as determined by the use of fluorogenic substrates for these enzymes. e) The bisphosphonate pamidronate (APD) caused a dose-dependent inhibition of resorption by equine OCLs. APD also dose-dependently reduced the number of OCLs present in MNCEP cultures after 7d incubation but paradoxically increased the resorption undertaken by similar cultures at certain concentrations. f) Calcitonin inhibited resorption by equine OCLs apparently by inducing cytoplasmic retraction. g) Resorption by OCLs was also inhibited by insulin and the proton-pump inhibitor, bafilomycin.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available