Profiling and authentication of human cell lines using short tandem repeat (STR) loci: Report from the National Cell Bank of Iran.
Assurance of cell line homogeneity and capability of cell contamination detection are among the most essential steps of cell based research. Due to high discriminatory efficiency, low cost and reliability, analysis of short tandem repeats (STR) has been introduced as a method of choice for human cell line authentication. In the present study 13 Combined DNA Index System (CODIS) based STRs along with the gender determination (Amelogenin) gene were utilized to establish a reproducible approach for the authentication of 100 human cell lines deposited in the National Cell Bank of Iran (NCBI), using the polymerase chain reaction (PCR) method. PCR products were subsequently analyzed by polyacrylamide gel electrophoresis (PAGE) and visualized by silver staining followed by gel documentation and software analysis. STR profiles obtained were compared with those of the American Type Culture Collection (ATCC) and the Japanese Collection of Research Bioresource (JCRB) as STR references. We detected 18.8% cross contamination among the NCBI human cell lines. To our knowledge, this is the first report of authentication of human cell lines using the 13 CODIS core STRs combined with Amelogenin.
Azari
Shahram
S
National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.
Ahmadi
Nahid
N
Tehrani
Mahmood Jeddi
MJ
Shokri
Fazel
F
D053523:Amelogenin / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D007492:Iran; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D014015:Tissue Banks / Q000592:standards
D053523:Amelogenin; D017931:DNA Primers