Differentiation of human osteoclasts from blood samples
Research type
Research Study
Full title
Differentiation of human osteoclasts from blood samples
IRAS ID
321094
Contact name
Caroline Gorvin
Contact email
Sponsor organisation
University of Birmingham
Duration of Study in the UK
4 years, 8 months, 31 days
Research summary
We will procure leukocyte cones (blood samples) from the NHSBT. Bone is a dynamic tissue that is remodelled throughout life by bone resorbing (i.e. bone eating) osteoclasts and bone forming osteoblasts, to adapt to physiological demands. These processes are impaired in osteoporosis and understanding how bone remodelling is regulated could improve anti-osteoporotic treatments. Our research aims to identify proteins that affect how human osteoclasts develop or how they destroy bone. By understanding how these proteins act in human osteoclasts, we could develop new drugs that reduce osteoclast activity and protect bone for the treatment of osteoporosis.
To perform these investigations, we need human osteoclasts which we can develop from human blood samples. We collect these samples (leukocyte cones) from the NHSBT. These samples have no identifying information for the patients. We then select blood monocytes using a specific marker (CD14). We seed these monocytes into tissue culture dishes and grow them in a nutrient mix comprising α-minimal essential medium (αMEM) with 10% fetal bovine serum (FBS), 1% penicillin/ streptomycin and 25 ng/mL macrophage colony-stimulating factor (MCSF). Media should be replaced every 2-3 days. On day 7-8 we replace this nutrient mix with a fresh mixture containing MCSF and receptor activator of nuclear κB (RANKL) (25 ng/mL). RANKL is used as it specifically activates the development of human osteoclast cells. We then perform assays to assess how different proteins activate or inhibit osteoclast function.
REC name
Wales REC 7
REC reference
23/WA/0063
Date of REC Opinion
20 Feb 2023
REC opinion
Further Information Favourable Opinion