军事医学2017，Vol.41Issue(3)：178-183,198,7.DOI:10.7644/j.issn.1674-9960.2017.03.005
识别多种种属来源间充质干细胞核酸适配体的筛选及鉴定
Selection and identification of aptamers for mesenchymal stem cells from different species
摘要
Abstract
Objective To screen an ssDNA aptamer for rabbit mesenchymal stem cells (MSCs) and to identify the ability of the aptamer to recognize MSCs of a variety of species origin.Methods MSCs were isolated from the thigh bone of immature rabbits and identified by induced osteogenic and adipogenic differentiation,respectively.Aptamers were screened by cell SELEX (systematic evolution of ligands by exponential enrichment) technique targeting to isolated MSCs.Enrichment of the 5th pool was evaluated through binding assay of FAM modified pool to MSCs by confocal microscopy.The enriched 5th pool was then cloned into pGE-T vector and the cloned sequences were determined randomly.The candidates were chosen based on primary sequence conservation and predicted secondary structure by RNA structure and MEME online analysis.Flow cytometry analysis was used to identify the aptamers binding to MSCs of rabbit, rat, and human origin.Results The isolated MSCs had the potential of osteogenic differentiation and adipogenic differentiation under certain conditions.Aptamer 5-1-12 from 5th enriched pool was characterized as MSCs recognizing aptamer binding to MSCs of rabbit, rat and human origin.Conclusion Aptamer 5-1-12 that recognizes MSCs of different species origin is obtained through live cell-SELEX.关键词
间充质干细胞/适配体/活细胞-SELEXKey words
mesenchymal stem cells/aptamer/live cell SELEX分类
医药卫生引用本文复制引用
郭晓华,李章华,汪伟,董洁,丁红梅,李少华,邵宁生..识别多种种属来源间充质干细胞核酸适配体的筛选及鉴定[J].军事医学,2017,41(3):178-183,198,7.基金项目
国家自然科学基金资助项目(81572846) （81572846）
湖北省自然科学基金重点资助项目(2015CFA079) （2015CFA079）
