Comparative Analysis of Protein Production by Human Chondrocyte Sheets Using a Multiplexed Aptamer-b

来源 :2016年国际组织工程与再生医学学会亚太会议 | 被引量 : 0次 | 上传用户:luomingasdf
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  [Objective] As we aim for regenerative treatment of knee cartilage using allogeneic chondrocyte sheets,we have examined chondrocytes obtained from polydactyly surgery as an alternative cell source.Secreted protein factors by chondrocyte sheets are likely to be significant contributors to cartilage regeneration; thus in this study,we compared the protein factors produced by chondrocyte sheets to estimate the sheets in vivo efficacy on cartilage regeneration.[Methods] Experiments were performed under the approval and guidance of the Clinical Research Review Committee of Tokai University School of Medicine.The chondrocytes were isolated from cartilaginous tissue from young polydactyly patients(PD: 7 patients; average age = 14.7 months)and patients undergoing surgery for total knee arthroplasty(TKA: 7 patients; average age = 76.1 years).The chondrocyte sheets were created using temperature-responsive culture dishes.The culture supernatants of chondrocyte sheets were analyzed using the SOMAscanTM: a multiplexed aptamer-based assay detecting 1129 proteins by slow off-rate modified aptamers(SOMAmers).[Results] Statistically significant differences(t-test: P-value < 0.01)between PD sheets and TKA sheets were observed in 211 of 1129 proteins.SOMAscanTM Contains 39 of 236 proteins in gene ontology(GO)term GO0051216: cartilage development.Among these 39 proteins,bone morphogenetic protein-10(BMP-10)and basic fibroblast growth factor(bFGF)were identified as differently produced proteins between PD and TKA sheets.[Discussion] Considering that TKA sheets are equivalent to adult chondrocyte sheets that demonstrated their efficacy in our recent clinical study,the difference between TKA sheets and PD sheets is important to consider.The contribution of BMP10 and bFGF to chondrocyte sheets efficacy on cartilage regeneration should be further studied for application of polydactyly chondrocytes as an alternative cell source.
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Introduction: We have studied the effects of chondrocyte sheets on the repair and regeneration of articular cartilage. With approval of the Ministry of Health, Labour and Welfare in Japan, we have per
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