Because of the risks that cell line contamination or misrepresentation pose during biomedical research, the NIH has developed guidelines for authenticating cells in order to receive funding and many journals now require confirmation before publication. The CWRU Genomics Core now offers comprehensive Cell Line Authentication Services using the Promega GenePrint 10 System. This 10-Locus short tandem repeat (STR) system is a fast, easy, and reliable way to profile human cell lines and confirm the identity.
Recommendations for when to authenticate a cell line:
- every time a new cell line is acquired or generated
- before freezing
- every two months that the cell is actively growing
- at any point that the performance of the line is inconsistent or yields unexpected results
- before publication
We offer a full service option for Cell Line Authentication to all CWRU users and affiliates. Starting with extracted DNA, the PCR is completed using the GenePrint 10 system and loaded onto the ABI 3730 DNA Analyzer. Raw data is analyzed with the GeneMapper software, using the given 2800M Control DNA as a positive control. The DSMZ database is used as a comparative tool to cross-reference STR profile queries against hundreds of human cell lines. The DSMZ is the most comprehensive biological resource center worldwide and is considered a highly respected and trusted site for cell line authentication information. A full report is released containing an STR profile table, an electropherogram profile, and a table with the raw data for each allele (size in base pairs, peak height, peak area). It also includes the profile comparison to the public database maintained by DSMZ.
"To standardize STR analysis for human cell line authentication, the American Tissue Culture Collection (ATCC) Standards Development Organization Workgroup published the ASN-0002-2011, which recommends the use of at least eight STR loci (TH01, TPOX, vWA, CSF1PO, D16S539, D7S820, D13S317 and D5S818) plus Amelogenin for gender identification." www.promega.com
| TH01 | D21S11 |
| D5S818 | D13S317 |
| D7S820 | D16S539 |
| CSF1PO | Amelogenin |
| vWA | TPOX |
Sample Submission Requirements
- Use a Nanodrop to calculate concentration and purity of extracted DNA. Normalize all samples to 10 ng/uL per Nanodrop. Purity should fall within the range of 1.8 - 2.0 260/280nm.
- Submit 25 uL of normalized DNA (10 ng/uL per Nanodrop) in a 1.5 mL tube, clearly labeled with sample name, user initials, and date.
- Fill out the Cell Line Authentication request form in iLab. Sample names listed should match the tubes submitted.