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Title: Identifying novel cell surface markers for bone marrow stem cell sub-sets
Author: Sutton , Catherine Anne
ISNI:       0000 0004 2721 0016
Awarding Body: University of Bristol
Current Institution: University of Bristol
Date of Award: 2012
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Osteoarthritis (OA) is a disease which affects hyaline cartilage within the joint and leads to severe pain for the patient. Chondrocytes within the cartilage of OA patients have a limited capability to repair lesions in the tissue and as a result, bone marrow stromal cells (BMSCs) which have the ability to differentiate into cartilage, are being investigated for cell- based therapies for OA. One restriction for the use of BMSCs in clinical trials is the rarity and heterogeneity of the starting population. It is therefore important to identify cell surface markers for BMSCs which produce a high quality cartilage matrix after many population doublings in vitro. Tissue engineered cartilage produced by BMSCs from OA patients at early and late passage after thawing was analysed and compared and it was found that the amount and quality of cartilage matrix was significantly reduced at late passage. To identify the most chondrogenic cells, individual BMSCs were sorted from five OA patients using flow cytometry, cultured for 20 population doublings and assessed for their cartilage tissue engineering capacity. BMSC clones which produced high quality tissue engineered cartilage were compared with BMSCs which produced a low quality cartilage using microarray analysis to find the genetic identities of the different populations. These markers were tested at the protein level using flow cytometric analysis and one protein, HLA DR was identified as a potential marker for poorly chondrogenic BMSCs. HLA DR positive cells were stably removed from the whole population and removal of these cells did not affect the quality of tissue engineered cartilage produced. Two protein markers for the most chondrogenic BMSCs were identified and investigated, FGFR-2 and ROR-2. There were no significant differences in the quality of tissue engineered cartilage produced by FGFR2 positive and negative populations. ROR-2 expression increased in culture as cells became confluent and cells cultured at a high cell density expressing increased ROR-2 had a trend to produce higher quality tissue engineered cartilage than those cultured at low density before chondrogenic analysis. These results highlight the heterogeneity in the cartilage producing capabilities of the BMSC population at the level of the individual cell and document the discovery of ROR-2 as a potential marker for a BMSC population which could be utilised in trials of BMSC based therapies for OA in the future.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available