人CD146+牙周膜干细胞的分选及其干细胞特性的鉴定

Isolation and identification of human CD146 positive periodontal ligament stem cells

目的:通过酶消化法及流式细胞分选技术分选CD146+牙周膜干细胞(Periodontal ligament cells,PDLCs),探讨其干细胞特性。方法:采用流式细胞技术从人牙周膜细胞中分选CD146+PDLCs,并对其进行免疫组化染色(角蛋白、波形丝蛋白及Stro-1),成骨、成脂诱导分化及克隆形成实验鉴定其干细胞特性。结果:CD146+PDLCs表达波形丝蛋白及Stro-1,具有成骨及成脂分化能力,体外培养能形成细胞克隆。结论:以CD146为分选指标的流式细胞分选技术可作为牙周膜干细胞筛选的方法。

Objectives： To isolate and culture CD146 positive periodontal ligament cells （CD146＋PDLCs） by using type I collagenase digestion and flow cytometric sorting, in order to verify the stem cell properties of CD146＋PDLCs. Methods： CD146＋PDLCs were isolated by flow activated cell sorter fi＇om human PDLCs. The stem cell properties of CD146＋PDLCs were tested by immunohistochemical staining, osteogenic/adipogenic induction and colony-forming assay. Results： CD146＋PDLCs, expressing mesenchymal stem cell surface marker Stro-1, were able to form calcium nodules/lipid droplets under osteogenic/adipogenic induction and were capable of forming colonies. Conclusions： CD 146＋PDLCs shared the similar properties with periodontal ligament stem cells, which providing a new method for periodontal ligament stem cell isolation.