Cell lines are essential in biomedical research, serving as models for various scientific experiments. However, issues related to cell line contamination, misidentification, and cross-contamination have raised concerns about the reliability & validity of research findings. To address these challenges, cell line authentication through DNA testing has become indispensable in ensuring research data’s accuracy and reproducibility. In India, the need for robust cell line authentication has grown with the increasing emphasis on cutting-edge research. This blog explores the significance of cell line authentication DNA test in India and how it helps to maintain research integrity and credibility.
Significance of Cell Line Authentication DNA Test
- Guaranteeing Scientific Integrity – Cell line authentication services in India ensure that the cell lines used in research are indeed what researchers believe them to be. Authenticating cell lines prevents errors arising from misidentification, cross-contamination, and genetic drift, thus ensuring the desired research outcomes.
- Avoiding Unreliable Data – Misidentified or contaminated cell lines can lead to erroneous data, leading researchers astray and wasting time & resources. Cell Line Authentication DNA Test mitigates this risk, giving researchers confidence in their experimental results.
- Reproducibility & Comparability – Authentic cell lines allow other researchers to replicate experiments and verify findings. Cell line authentication ensures that studies can be reproduced accurately and reliably, promoting the advancement of scientific knowledge.
- Ethical Considerations – When cell lines are misidentified, it can raise ethical concerns. Using the wrong cell type in experiments may have unintended consequences for research outcomes. By verifying the identity of cell lines, researchers can ensure ethical and accurate research practices.
Human Cell Line Authentication Methodology – By STR Profiling Technique
Cell line authentication involves the use of various techniques to verify the identity and purity of cell lines. The most commonly employed method is short tandem repeat (STR) profiling, which analyzes specific regions of the DNAs of the cell lines of interest to create a unique genetic profile. There are two options for providing the sample: extracted DNA or cell pellet.
Extracted DNA Method:
- Collect samples in labeled 1.5-2.0 ml microcentrifuge tubes.
- Extract and purify the DNA and then elute it in Low TE or Nucleolus Free Water. Ensure a concentration of 10-100ng/µl*. Quantify DNA concentration with a qubit fluorometer (not a Nanodrop spectrophotometer) and use 10-20 µl volume per tube.
- Seal the tubes properly with paraffin to avoid leakage.
- Freeze the tubes at -20°C and pack them in a zip bag with ice packs for shipping.
Cell Pellet Method:
- Collect the sample in labeled 1.5-2.0 ml microcentrifuge tubes.
- Protocol for attached cells – Trypsinize the cell culture for attached cells and centrifuge at 125 x g. Resuspend the pellet in PBS** to achieve a concentration of 1.6 x 106 cells/ml.
- Protocol for suspension cells – For suspension cells, collect and count cells to achieve a concentration of 1.0 x 106 cells/ml. Dilute with PBS if highly concentrated or centrifuge and resuspend in PBS if already diluted.
- Use 1.5-2.0 ml microcentrifuge tubes for cell suspension.
*Ng/ µl – nanogram per microliter
**PBS (Phosphate Buffered Saline) – It is a non-toxic solution. Unlike water, PBS prevents cells from rupturing or shrivelling due to osmosis.
- Conduct a PCR reaction with fluorescent primers recognizing different loci.
- Add internal standards for size after PCR completion and separate DNA by size using capillary gel electrophoresis.
- Analyze size using GeneMapper ID-X (GMIDX) software, which compares DNA fragment sizes with internal standards.
- Perform STR genotyping by converting amplicon sizes into alleles using the allelic ladder.
STR profiling helps ensure the accurate identification and validation of human cell lines, essential for maintaining ethical research practices and reliable research outcomes. Furthermore, when submitting more than one sample, please ensure to complete a separate cell line authentication sample submission form for each sample.
Why DNA Forensics Laboratory Stands Out for Cell Line Authentication Services in India?
DNA Forensics Laboratory Pvt. Ltd. is among a leading provider of accurate & reliable cell line authentication services in India. Our services cover both human and mouse cell line authentication, enabling us to identify misidentified, genetically drifted, and cross-contaminated cell lines effectively.
We proudly collaborate with prestigious research institutes, pharmaceutical companies, and biotechnology firms. Our cell line authentication DNA test reports include an easy-to-understand STR allele table, supporting electropherograms at each locus, and a detailed interpretation of the results. Additionally, we compare the cell’s profile against our comprehensive database.
Furthermore, we are the only company that offers legal DNA tests in India. With 400+ collection centers nationwide and internationally, you can conveniently visit your nearest location to provide your cell lines.
All in all, our commitment to quality, expertise, and timely delivery service has made us the preferred choice for researchers, scientists, and professionals in need of reliable cell line authentication services. For further inquiries about cell line authentication DNA Tests in India, reach out to us at +91 8010177771 or chat with us on WhatsApp at +91 9213177771 to schedule an appointment.