title: Yield optimisation and molecular characterisation of uncultured CD271+ mesenchymal stem cells in the Reamer Irrigator Aspirator waste bag.
abstract: Bone reconstruction requires the use of autografts from patients' iliac crest (IC); for large-volume defects bone void fillers and autologous mesenchymal stem cells (MSCs) are often added. The Reamer/Irrigator/Aspirator (RIA) device provides the means of harvesting large amounts of autograft and additionally yields a waste bag containing MSCs, which is currently discarded. The aim of this study was to enumerate and characterise native MSCs from RIA waste bag and compare them to 'gold-standard' donor-matched MSCs from IC bone marrow (BM). IC-BM from age matched trauma patients was used as control. In RIA waste bags the median MSC yield established using a colony-forming fibroblast assay was 314333 (range 5 x 104-1.4 x 106), equivalent to approximately one litre of IC-BM aspirate. CD271+ cells were present at high levels in RIA waste bags, had MSC surface phenotype (CD90+CD73+CD105+CD34>sup>-CD61-CD19-CD31-CD33-) and expressed genes associated with multipotentiality, osteogenesis, adipogenesis and angiogenic support. RIA- CD271+ MSCs were transcriptionally similar to donor-matched IC-CD271+ MSCs (76 % transcripts); with the majority of bone-related and Wnt pathway molecules being expressed at comparable levels. Lower-level expression of MCAM/CD146 and 5/13 hypoxia-related molecules was found in RIA-CD271+ MSCs, potentially reflecting their native residence in a more hypoxic environment of the endosteum and bone cortex. These data suggest that long bones contain very large numbers of MSCs, transcriptionally-similar to IC-BM MSCs; they can be procured by reaming using the RIA device and used, following concentration, as autologous and potentially allogeneic bone repair therapy.
authors: Academic Department of Trauma & Orthopaedic Surgery, A Floor, Clarendon Wing, Leeds General Infirmary, Great George Street, Leeds LS1 3EX, UK.pgiannoudi@aol.com.
authors: Sarah M
authors: SM
authors: Churchman
authors: Dimitrios
authors: D
authors: Kouroupis
authors: Sally A
authors: SA
authors: Boxall
authors: Tarek
authors: T
authors: Roshdy
authors: Hiang B
authors: HB
authors: Tan
authors: Dennis
authors: D
authors: McGonagle
authors: Peter V
authors: PV
authors: Giannoudis
authors: Elena A
authors: EA
authors: Jones
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000295:instrumentation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008875:Middle Aged; D007507:Therapeutic Irrigation / Q000295:instrumentation; D020858:Tissue and Organ Harvesting / Q000295:instrumentation / Q000379:methods*; D014182:Transplantation, Autologous / Q000379:methods
chemical_list: D000943:Antigens, Differentiation