Browsing by Author "Al-Jezani, Nedaa"

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    Cell Surface Receptors Expression Profile Of Human Synovial Mesenchymal Stem Cells In-Situ And In-Vitro From Normal And Osteoarthritic Knee Joints
    (2016) Al-Jezani, Nedaa; Krawetz, Roman; Rancourt, Derrick; Duncan, Neil
    Osteoarthritis is a chronic disease that results in the degeneration of the articular cartilage, eventually affecting the whole joint. Due to the lack of regenerative capacity in the articular cartilage, synovial mesenchymal stem cells, having chondrogenic capacity and derived from the synovium (SMSCs) are an attractive source for articular cartilage regeneration. However, it is important to identify the optimal SMSCs that have tri-lineage potential, which will help the regeneration of the whole joint. Unfortunately, MSC populations are phenotypically heterogeneous, in terms of their cell surface expression. In this project, SMSCs derived from OA joint with the optimal differentiation capacity display a marker profile of CD90+, CD44+, and CD73+. However, the same profile was expressed on clones that didn’t present with multi-potential differentiation capacity. Therefore, additional markers are necessary to accurately purify SMSCs that would may have increased regenerative capacity for use in cell therapies for patients with OA.
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    Isolation and Characterization of an Adult Stem Cell Population from Human Epidural Fat
    (2019-03-17) Al-Jezani, Nedaa; Cho, Roger; Masson, Anand O.; Lenehan, Brian; Krawetz, Roman; Lyons, Frank G.
    Study Design. Isolation and characterization of human epidural fat (HEF) stem/progenitor cells. Objective. To identify a progenitor population within HEF and to determine if they meet the minimal criteria of a mesenchymal stem cell (MSC). Summary of Background Data. The biological function, if any, has yet to be determined for HEF. The presence of MSCs within HEF may indicate a regenerative potential within the HEF. Methods. HEF was isolated from 10 patients during elective spinal surgery. HEF cells were differentiated along osteo-, adipo-, and chondrogenic lineages, with differentiation analyzed via qPCR and histology. The cell surface receptor profile of HEF cells was examined by flow cytometry. HEF cells were also assayed through the collagen contraction assay. Prx1CreERT2GFP:R26RTdTomato MSC lineage-tracking mice were employed to identify EF MSCs in vivo. Results. HEF cell lines were obtained from all 10 patients in the study. Cells from 2/10 patients demonstrated full MSC potential, while cells from 6/10 patients demonstrated progenitor potential; 2/10 patients presented with cells that retained only adipogenic potential. HEF cells demonstrated MSC surface marker expression. All patient cell lines contracted collagen gels. A Prx1-positive population in mouse epidural fat that appeared to contribute to the dura of the spinal cord was observed in vivo. Conclusions. MSC and progenitor populations are present within HEF. MSCs were not identified in all patients examined in the current study. Furthermore, all patient lines demonstrated collagen contraction capacity, suggesting either a contaminating activated fibroblast population or HEF MSCs/progenitors also demonstrating a fibroblast-like phenotype. In vivo analysis suggests that these cell populations may contribute to the dura. Overall, these results suggest that cells within epidural fat may play a biological role within the local environment above providing a mechanical buffer.
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    Multiple mesenchymal progenitor cell subtypes with distinct functional potential are present within the intimal layer of the hip synovium
    (2019-03-25) Affan, Asmaa; Al-Jezani, Nedaa; Railton, Pamela; Powell, James N; Krawetz, Roman J
    Abstract Background The synovial membrane adjacent to the articular cartilage is home to synovial mesenchymal progenitor cell (sMPC) populations that have the ability to undergo chondrogenesis. While it has been hypothesized that multiple subtypes of stem and progenitor cells exist in vivo, there is little evidence supporting this hypothesis in human tissues. Furthermore, in most of the published literature on this topic, the cells are cultured before derivation of clonal populations. This gap in the literature makes it difficult to determine if there are distinct MPC subtypes in human synovial tissues, and if so, if these sMPCs express any markers in vivo/in situ that provide information in regards to the function of specific MPC subtypes (e.g. cells with increased chondrogenic capacity)? Therefore, the current study was undertaken to determine if any of the classical MPC cell surface markers provide insight into the differentiation capacity of sMPCs. Methods Clonal populations of sMPCs were derived from a cohort of patients with hip osteoarthritis (OA) and patients at high risk to develop OA using indexed cell sorting. Tri-differentiation potential and cell surface receptor expression of the resultant clones was determined. Results A number of clones with distinct differentiation potential were derived from this cohort, yet the most common cell surface marker profile on MPCs (in situ) that demonstrated chondrogenic potential was determined to be CD90+/CD44+/CD73+. A validation cohort was employed to isolate cells with only this cell surface profile. Isolating cells directly from human synovial tissue with these three markers alone, did not enrich for cells with chondrogenic capacity. Conclusions Therefore, additional markers are required to further discriminate the heterogeneous subtypes of MPCs and identify sMPCs with functional properties that are believed to be advantageous for clinical application.