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Strand displacement amplification for ultrasensitive detection of human pluripotent stem cells
- Source :
- Analytica Chimica Acta. 881:124-130
- Publication Year :
- 2015
- Publisher :
- Elsevier BV, 2015.
-
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.
- Subjects :
- Pluripotent Stem Cells
Stage-Specific Embryonic Antigens
Cell Culture Techniques
DNA, Single-Stranded
Metal Nanoparticles
Biosensing Techniques
Sensitivity and Specificity
Biochemistry
Regenerative medicine
Analytical Chemistry
Humans
Environmental Chemistry
Antigens, Tumor-Associated, Carbohydrate
Induced pluripotent stem cell
Spectroscopy
Chemistry
Multiple displacement amplification
Equipment Design
Nucleic acid amplification technique
Embryonic stem cell
Molecular biology
Cell culture
Gold
Stem cell
Antibodies, Immobilized
Nucleic Acid Amplification Techniques
Subjects
Details
- ISSN :
- 00032670
- Volume :
- 881
- Database :
- OpenAIRE
- Journal :
- Analytica Chimica Acta
- Accession number :
- edsair.doi.dedup.....6a2d27d21f175602acc8c42bffb72606