CLA-Power of small molecular stretched STR for Authentication of Human Cell Lines
Authentication or purity check of parental cell lines are utmost important to investigate and interrogate, purity of cultures to be propagated for the commercial and research application. Any contamination of cell line (s) at any stage of processing or cross contamination while new cell line development or lab contamination or storage or seed culture limit final resources, financial burden as well final production outcome. Most of international publication refer CLA for any human cell related work to be published or commercialized for the product development including cancer discoveries to cancer therapeutics and other related disciplines.
Short Tandem Repeat (STR) DNA profiling of human cell lines, is one of the versatile tools for cell culture management. However, other methodologies are available like genotyping by SNPs, karyotyping, PCR specific primers, STR-Based Cell lines authentication so far one of the promising and cost effective tools. STR based primers and fluorescent probes allow more sensitive detection of any alternation or genetic drift over the course of time, primarily due to shorter repetitive sequences of 3-7 bases, separated by no of copies spanning within PCR amplified region.
Sanger sequencing or genetic analyzer is one the tools using automated capillary electrophoresis for the size selection of allelic distribution with different sizes determine the marker for cell line profile study. Based on the selective size of Allele, separated over the Genetic Analyzer assigned specific market of each STR.
Fig: STR analysis work flow (Curtsey of Promega corporation)
The Major advantage of STR based Cell authentication over other methodologies will be
- Amplified STR regions are shorter, likely to me more stable and less subject to degradation
- Sample size and input DNA not limited to be subjected for authentication
- Major advantage of power and comparative profiling with other methods
- Power of multiplexing with ease of diverse primers and fluorescent probes in single test
- Over all processing and result interpretation is very less.
Eurofins Genomics India Pvt. Ltd offer CLA end to end including DNA isolation to raw data generation to check purity of cell line to save the risk of any commercial cell therapeutics production.