Large‐scale extraction and characterization of CD271+ multipotential stromal cells from trabecular bone in health and osteoarthritis: Implications for bone regeneration strategies based on uncultured or minimally cultured multipotential stromal cells

Abstract

Objective. To test the hypothesis that CD45 low CD271؉ bone marrow multipotential stromal cells (MSCs) are abundant in the trabecular bone niche and to explore their functional "fitness" in health and osteoarthritis (OA).Methods. Following enzymatic extraction, MSC release was evaluated using colony-forming unitfibroblast (CFU-F) and colony-forming unit-osteoblast assays, flow cytometry, and confocal microscopy. CD45 low CD271؉ cells isolated by fluorescence-activated cell sorting were enumerated and expanded under standard and clonal conditions. Their proliferative and osteogenic potencies were assessed in relation to donor age and compared with those of aspirated CD45 low CD271؉ cells. In vitro and in vivo MSC "aging" was measured using quantitative polymerase chain reaction-based telomere length analysis, and standard differentiation assays were utilized to demonstrate multipotentiality.Results. Conclusion. Our findings show that CD45 low CD271؉ MSCs are abundant in the trabecular bone cavity and indistinguishable from aspirated CD45 low CD271؉ MSCs. In OA they display agingrelated loss of proliferation but no gross osteogenic abnormality. These findings offer new opportunities for direct study of MSCs in musculoskeletal diseases without the requirement for culture expansion. They are also relevant for direct therapeutic exploitation of prospectively isolated, minimally cultured MSCs in trauma and OA.

Interleukin-22 drives the proliferation, migration and osteogenic differentiation of mesenchymal stem cells: a novel cytokine that could contribute to new bone formation in spondyloarthropathies

“…MSC functionality was confirmed for cells used in subsequent experiments using in vitro tri-lineage MSC differentiation assays in representative samples (n=6) as previously described [12] (data not shown).…”

Mesenchymal Stem Cell Alterations in Bone Marrow Lesions in Patients With Hip Osteoarthritis

“…Our previous analysis of CD45−CD271+ cells sorted from whole OA femoral heads did not reveal any significant signs of premature aging or gross osteogenic abnormality compared with control bone 11. In the current study, we carefully excised BML and non‐BML areas of a femoral head, as segregated using MRI, and were able to detect subtle differences in MSC features within the same affected joint but in relation to the amount of tissue damage.…”

Yield optimisation and molecular characterisation of uncultured CD271+ mesenchymal stem cells in the reamer irrigator aspirator waste bag

“…+ cells were first pre-enriched with Anti-Fibroblast Microbeads (Miltenyi), as described previously Jones et al, 2010). The MSC population: CD45 -/ low CD271 + , together with the control haematopoietic lineage cell (HLC) population; CD45 + CD271 -, was purified by cell sorting, using CD45-FITC (Dako, Stockport, UK), CD271-PE (BD Biosciences, Oxford, UK) and 7-AAD (Sigma, Gillingham, UK), the latter used to eliminate dead/dying cells, on a MoFlo cell sorter (Dako, Ely, UK), directly into lysis buffer (Norgen Biotek, Thorold, Canada).…”