JUL 02, 2020 12:15 PM SGT

Authenticating human cells: Knowing exactly what is in your test tube

Speaker

Abstract

The study of human development and diseases relies on analysis of samples obtained and manipulated ex vivo. It is critical in such studies to know the provenance and confirm that the identity of these materials is correct. Analysis of highly variable short tandem repeats (STRs), provides a simple, inexpensive and highly specific genetic “fingerprint” of a human sample.  In this study, we describe a complete workflow for cell line authentication, chimeric receptor T-cell (CAR-T) and induced pluripotent stem cell (iPSC) sample matching.  Analysis of serial dilutions of purified genomic DNA from isolated human cell lines identified the correct alleles from as little as 0.1ng of purified DNA.  Samples immobilized onto Copan NUCLEIC Cards were correctly identified when a suspension of 5-10 x 105 cells were dried onto the cards.  Contaminating HeLa alleles could be detected in mixture when they were present in as little as 1% of genomic DNA from this population.  Blinded samples of donor and iPSCs, as well as manipulated CAR-T cells, were correctly matched in every case tested. Together, these results describe a facile and consolidated workflow for establishing the provenance, authenticity and identity of ex vivo-obtained human samples.


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