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目的筛选能识别多种属来源的间充质干细胞(MSC)的核酸适配体,并对其进行鉴定。方法从幼兔股骨骨片分离MSC并进行成骨、成脂诱导分化鉴定。以分离得到的兔源性MSC为筛选靶标,利用细胞SELEX技术筛选核酸适配体。将第5轮富集文库进行克隆、测序,利用RNA structure、MEME软件分析获得候选序列,利用流式细胞仪鉴定候选适配体与兔、大鼠、人源性MSC的结合。结果从兔股骨骨片分离得到的MSC可诱导向成骨、成脂分化。经过5轮SELEX筛选获得兔MSC富集文库。候选适配体5-1-12不仅结合兔MSC,也识别大鼠和人源性MSC。结论活细胞SELEX筛选获得与多种属来源MSC结合的核酸适配体5-1-12。
Objective To screen and identify nucleic aptamers that recognize mesenchymal stem cells (MSCs) of various genera. Methods MSCs were isolated from the femoral bone fragments of rabbits and osteogenic and adipogenic differentiation was performed. Using isolated rabbit-derived MSCs as screening targets, SELEX technology was used to screen aptamers. The fifth round of enrichment library was cloned and sequenced. The candidate sequences were obtained by RNA structure and MEME software analysis. The binding of candidate aptamers to rabbit, rat and human MSC were identified by flow cytometry. Results MSC isolated from rabbit femoral bone chips could induce osteogenic and adipogenic differentiation. Rabbit MSC enrichment library was obtained after 5 rounds of SELEX screening. Candidate aptamers 5-1-12 not only bind rabbit MSCs, but also rat and human-derived MSCs. Conclusion SELEX screening of living cells obtained aptamers 5-1-12 that bind to MSC of various genera.