CAJ | 학술논문

目的研究体外培养的人牙周膜细胞群(hPDLP)向成骨和成脂方向分化的潜能,为牙周组织工程提供可靠的种子细胞来源.方法组织块法分离培养人牙周膜细胞群,流式细胞术检测间充质干细胞标记CD146和STRO-1的表达;利用茜素红、油红O染色、免疫组化以及反转录聚合酶链反应(RT-PCR)等检测hPDLP的多向分化标志.结果第1代hPDLP的CD146和STRO-1阳性率分别是27.20%±3.98%和4.23%±4.08%;经矿化诱导可以形成矿化结节,有钙盐沉积;经成脂诱导可见特异性脂滴形成,特异性转录因子过氧化物酶体激活物增生受体2(PPARγ2)和脂蛋白脂酶(LPL)表达上调.第8代hPDLP相对第1代的矿化能力没有差异,但成脂方向分化潜能减弱.结论体外培养的hPDLP具有向成骨和成脂样细胞分化潜能,第1～3代细胞群明显具有牙周膜干细胞的多向分化潜能优势.
목적 연구체외배양적인아주막세포군(hPDLP)향성골화성지방향분화적잠능,위아주조직공정제공가고적충자세포래원.방법 조직괴법분리배양인아주막세포군,류식세포술검측간충질간세포표기CD146화STRO-1적표체;이용천소홍、유홍O염색、면역조화이급반전록취합매련반응(RT-PCR)등검측hPDLP적다향분화표지.결과 제1대hPDLP적CD146화STRO-1양성솔분별시27.20%±3.98%화4.23%±4.08%;경광화유도가이형성광화결절,유개염침적;경성지유도가견특이성지적형성,특이성전록인자과양화물매체격활물증생수체2(PPARγ2)화지단백지매(LPL)표체상조.제8대hPDLP상대제1대적광화능력몰유차이,단성지방향분화잠능감약.결론 체외배양적hPDLP구유향성골화성지양세포분화잠능,제1～3대세포군명현구유아주막간세포적다향분화잠능우세.
Objective To explore the multi-differentiated capability of human periodontal ligament cell population (hPDLP), and provide a theoretical basis for the periodontal regeneration by tissue engineering technique. Methods hPDLP was cultured from periodontium of human tooth by the outgrowth method. STRO-1 and CD 146 expression were investigated by flow cytometry. hPDLP was induced to odontogenic/osteogenic-like and adipogenic-like cell. The multilineage differentiation capacities of hPDLP were evaluated by alizarin red stain, oil red 0 stain, anti-CD146 and STRO-1 immunocytochemistry, and reverse-transcriptase polymerase chain reaction analysis. Results hPDLP was isolated from human periodontium and most of the cells retained their fibroblastic spindle shape. hPDLP can be induced into osteoblast-like cells and adipocyte-like cells, and calcium deposition and lipid droplets were detected perspectively. And the eighth generation of hPDLP had weaker potential into adipocyte-like cells than the first passage, however, there was no difference to the aspect of calcification ability between the two passages. Conclusion hPDLP cultured in vitro can differentiate into adipocytes and osteoblasts, and the first to third passage cells may have the predominance of differentiation potential.