title: Brain tumor cell line authentication, an efficient alternative to capillary electrophoresis by using a microfluidics-based system.
abstract: BACKGROUND
  The current method for cell line authentication is genotyping based on short tandem repeat (STR)-PCR involving coamplification of a panel of STR loci by multiplex PCR and downstream fragment length analysis (FLA), usually performed by capillary electrophoresis. FLA by capillary electrophoresis is time-consuming and can be expensive, as the facilities are generally not accessible for many research laboratories.
  
  
  METHODS
  In the present study, a microfluidic electrophoresis system, the Agilent 2100 Bioanalyzer, was used to analyze the STR-PCR fragments from 10 human genomic loci of a number of human cell lines, including 6 gliomas, 1 astrocyte, 1 primary lung cancer, 1 lung brain metastatic cancer, and 1 rhabdomyosarcoma; and this was compared with the standard method, that is, capillary electrophoresis, using the Applied Biosystems 3130xl Genetic Analyzer.
  
  
  RESULTS
  The microfluidic electrophoresis method produced highly reproducible results with good sensitivity in sizing of multiple PCR fragments, and each cell line demonstrated a unique DNA profile. Furthermore, DNA fingerprinting of samples from 5 different passage numbers of the same cell line showed excellent reproducibility when FLA was performed with the Bioanalyzer, indicating that no cross-contamination had occurred during the culture period.
  
  
  CONCLUSIONS
  This novel application provides a straightforward and cost-effective alternative to STR-based cell line authentication. In addition, this application would be of great value for cell bank repositories to maintain and distribute precious cell lines.
authors: Corresponding Author: Dr. Qian An, MD, PhD, School of Pharmacy and Biomedical Sciences, University of Portsmouth, St Michael's Building, White Swan Road, Portsmouth PO1 2DT, UK. qian.an@port.ac.uk.
authors: Qian
authors: Q
authors: An
authors: Helen L
authors: HL
authors: Fillmore
authors: Mikaella
authors: M
authors: Vouri
authors: Geoffrey J
authors: GJ
authors: Pilkington
mesh_terms: D001932:Brain Neoplasms / Q000145:classification / Q000235:genetics*; D016172:DNA Fingerprinting / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005838:Genotype; D006801:Humans; D044085:Microfluidics / Q000379:methods*; D018895:Microsatellite Repeats / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D014407:Tumor Cells, Cultured
keywords: DNA fingerprinting; STR profiling; brain tumor cell authentication; microfluidics