RESUMO
Use of D NA markers in research that are used in law enforcement risks undermining public trust and participation.
Assuntos
Autenticação de Linhagem Celular , Genética Forense , Aplicação da Lei , Análise de Sequência de DNA , Manejo de Espécimes , Confiança , Autenticação de Linhagem Celular/ética , Autenticação de Linhagem Celular/normas , Participação da Comunidade , Genética Forense/ética , Marcadores Genéticos , Humanos , Aplicação da Lei/ética , Análise de Sequência de DNA/ética , Manejo de Espécimes/éticaRESUMO
Colorectal and glioblastoma cancer stem-like cells (CSCs) are essential for translational research. Cell line authentication by short tandem repeat (STR) profiling ensures reproducibility of results in oncology research. This technique enables to identify mislabeling or cross-contamination of cell lines. In our study, we provide a reference dataset for a panel of colorectal and glioblastoma CSCs that allows authentication. Each cell line was entered into the cell Line Integrated Molecular Authentication database 2.1 to be compared to the STR profiles of 4485 tumor cell lines. This article also provides clinical data of patients from whom CSCs arose and data on the parent tumor stage and mutations. STR profiles and information of our CSCs are also available in the Cellosaurus database (ExPASy) as identified by unique research resource identifier codes.
Assuntos
Autenticação de Linhagem Celular/métodos , Autenticação de Linhagem Celular/normas , Linhagem Celular Tumoral , Repetições de Microssatélites , Células-Tronco Neoplásicas , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias Colorretais/genética , Conjuntos de Dados como Assunto , Feminino , Perfilação da Expressão Gênica/métodos , Perfilação da Expressão Gênica/normas , Glioblastoma/genética , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Cross-contamination of cell lines is a highly relevant and pervasive problem. The analysis of short tandem repeats (STR) is a simple and commercially available technique to authenticate cell lines for more than two decades. At present, STR multiple amplification kits have been developed up to 21 loci while the current STR databases only provide 9-loci STR profiles. Here, we compared the advantages of 21-loci STR methodology using the same algorithm as 9-loci method. The 21-loci method reduced the uncertainty ratio for authentications by 97.5% relative to the 9-loci method and exclude effectively false positive. We show that the additional 12 loci helped to greatly reduce sample-site marker specificity arising from genetic isolation and the occurrence of null alleles, suggesting that inclusion of additional loci in these databases will ultimately improve the efficiency and accuracy of authentication of cell lines. Taken together, we demonstrate the utility of a 21-loci method in human cells, providing a novel marker panel for use as a valuable alternative to 9-loci analyses to minimize cell line authentication errors and reduce costs due to erroneous experiments.
Assuntos
Autenticação de Linhagem Celular/métodos , Repetições de Microssatélites , Linhagem Celular , Autenticação de Linhagem Celular/normas , Linhagem Celular Tumoral , Loci Gênicos , Marcadores Genéticos , Humanos , Tipagem Molecular/métodos , Tipagem Molecular/normasRESUMO
Cell line authentication is important for credibility concern and scientific reproducibility. Authenticated cancer cell lines retain the properties of the cancers of origin and serve valuable resources for medical research. Experimental results commonly will be validated in more than one cell line to avoid specific cell line effect not generalizable to the disease on the whole. The use of appropriate and verified cell lines would therefore be very important in preclinical studies of translational research, bridging basic research to clinics.
