Recently,gingival margin-derived stem/progenitor cells isolated via STRO-1/magnetic activated cell sorting（MACS） showed remarkable periodontal regenerative potential in vivo.As a second-stage investigation,the prese...Recently,gingival margin-derived stem/progenitor cells isolated via STRO-1/magnetic activated cell sorting（MACS） showed remarkable periodontal regenerative potential in vivo.As a second-stage investigation,the present study＇s aim was to perform in vitro characterisation and comparison of the stem/progenitor cell characteristics of sorted STRO-1-positive（MACS~＋） and STRO-1-negative（MACS^-） cell populations from the human free gingival margin.Cells were isolated from the free gingiva using a minimally invasive technique and were magnetically sorted using anti-STRO-1 antibodies.Subsequently,the MACS~＋ and MACS^- cell fractions were characterized by flow cytometry for expression of CD14,CD34,CD45,CD73,CD90,CD105,CD146/MUC18 and STRO-1.Colony-forming unit（CFU） and multilineage differentiation potential were assayed for both cell fractions.Mineralisation marker expression was examined using real-time polymerase chain reaction（PCR）.MACS~＋ and MACS- cell fractions showed plastic adherence.MACS~＋ cells,in contrast to MACS- cells,showed all of the predefined mesenchymal stem/progenitor cell characteristics and a significantly higher number of CFUs（P〈0.01）.More than 95%of MACS~＋ cells expressed CD105,CD90 and CD73;lacked the haematopoietic markers CD45,CD34 and CD14,and expressed STRO-1 and CD146/MUC18.MACS- cells showed a different surface marker expression profile,with almost no expression of CD14 or STRO-1,and more than 95%of these cells expressed CD73,CD90 and CD146/MUC18,as well as the haematopoietic markers CD34 and CD45 and CD105.MACS~＋ cells could be differentiated along osteoblastic,adipocytic and chondroblastic lineages.In contrast,MACS- cells demonstrated slight osteogenic potential.Unstimulated MACS~＋ cells showed significantly higher expression of collagen I（P〈0.05） and collagen III（P〈0.01）,whereas MACS^- cells demonstrated higher expression of osteonectin（P〈0.05;MannWhitney）.The present study is the first to compare gingival MACS~＋ and MACS- cell populations demonstrating that MACS~＋ cells,in contrast to MACS- cells,harbour stem/progenitor cell characteristics.This study also validates the effectiveness of the STRO-l/MACS~＋technique for the isolation of gingival stem/progenitor cells.Human free gingival margin-derived STRO-1/MACS~＋ cells are a unique renewable source of multipotent stem/progenitor cells.展开更多
基金supported in part by a scholarship from the GermanAcademic-Exchange-Service(DAAD)
文摘Recently,gingival margin-derived stem/progenitor cells isolated via STRO-1/magnetic activated cell sorting（MACS） showed remarkable periodontal regenerative potential in vivo.As a second-stage investigation,the present study＇s aim was to perform in vitro characterisation and comparison of the stem/progenitor cell characteristics of sorted STRO-1-positive（MACS~＋） and STRO-1-negative（MACS^-） cell populations from the human free gingival margin.Cells were isolated from the free gingiva using a minimally invasive technique and were magnetically sorted using anti-STRO-1 antibodies.Subsequently,the MACS~＋ and MACS^- cell fractions were characterized by flow cytometry for expression of CD14,CD34,CD45,CD73,CD90,CD105,CD146/MUC18 and STRO-1.Colony-forming unit（CFU） and multilineage differentiation potential were assayed for both cell fractions.Mineralisation marker expression was examined using real-time polymerase chain reaction（PCR）.MACS~＋ and MACS- cell fractions showed plastic adherence.MACS~＋ cells,in contrast to MACS- cells,showed all of the predefined mesenchymal stem/progenitor cell characteristics and a significantly higher number of CFUs（P〈0.01）.More than 95%of MACS~＋ cells expressed CD105,CD90 and CD73;lacked the haematopoietic markers CD45,CD34 and CD14,and expressed STRO-1 and CD146/MUC18.MACS- cells showed a different surface marker expression profile,with almost no expression of CD14 or STRO-1,and more than 95%of these cells expressed CD73,CD90 and CD146/MUC18,as well as the haematopoietic markers CD34 and CD45 and CD105.MACS~＋ cells could be differentiated along osteoblastic,adipocytic and chondroblastic lineages.In contrast,MACS- cells demonstrated slight osteogenic potential.Unstimulated MACS~＋ cells showed significantly higher expression of collagen I（P〈0.05） and collagen III（P〈0.01）,whereas MACS^- cells demonstrated higher expression of osteonectin（P〈0.05;MannWhitney）.The present study is the first to compare gingival MACS~＋ and MACS- cell populations demonstrating that MACS~＋ cells,in contrast to MACS- cells,harbour stem/progenitor cell characteristics.This study also validates the effectiveness of the STRO-l/MACS~＋technique for the isolation of gingival stem/progenitor cells.Human free gingival margin-derived STRO-1/MACS~＋ cells are a unique renewable source of multipotent stem/progenitor cells.
