title: Strand displacement amplification for ultrasensitive detection of human pluripotent stem cells.
abstract: Human pluripotent stem cells (hPSCs), such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), provide a powerful model system for studies of cellular identity and early mammalian development, which hold great promise for regenerative medicine. It is necessary to develop a convenient method to discriminate hPSCs from other cells in clinics and basic research. Herein, a simple and reliable biosensor for stem cell detection was established. In this biosensor system, stage-specific embryonic antigen-3 (SSEA-3) and stage-specific embryonic antigen-4 (SSEA-4) were used to mark human pluripotent stem cells (hPSCs). Antibody specific for SSEA-3 was coated onto magnetic beads for hPSCs enrichment, and antibody specific for SSEA-4 was conjugated with carboxyl-modified tDNA sequence which was used as template for strand displacement amplification (SDA). The amplified single strand DNA (ssDNA) was detected with a lateral flow biosensor (LFB). This biosensor is capable of detecting a minimum of 19 human embryonic stem cells by a strip reader and 100 human embryonic stem cells by the naked eye within 80min. This approach has also shown excellent specificity to distinguish hPSCs from other types of cells, showing that it is promising for specific and handy detection of human pluripotent stem cells.
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wu
authors.affiliation: Yueyang Institute for Food and Drug Control, Yueyang 430198, China.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Mao
authors.affiliation: Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Shiming
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Xuewen
authors.initials: X
authors.lastname: Lu
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China. Electronic address: liangxg@ouc.edu.cn.
authors.forename: Xingguo
authors.initials: X
authors.lastname: Liang
authors.affiliation: Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China. Electronic address: zeng6@yahoo.com.
authors.forename: Lingwen
authors.initials: L
authors.lastname: Zeng
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D018929:Cell Culture Techniques; D004277:DNA, Single-Stranded / Q000032:analysis; D004867:Equipment Design; D006046:Gold / Q000737:chemistry; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000276:immunology; D012680:Sensitivity and Specificity; D055746:Stage-Specific Embryonic Antigens / Q000737:chemistry / Q000276:immunology
chemical_list: D055910:Antibodies, Immobilized; D015295:Antigens, Tumor-Associated, Carbohydrate; D004277:DNA, Single-Stranded; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3; C066584:stage-specific embryonic antigen-4; D006046:Gold
keywords: Human pluripotent stem cells; Lateral flow biosensor; Strand displacement amplification