High purity and integrity RNA from human cell lines stored in liquid nitrogen for over 20 years

Biobanks are a key resource for obtaining human cell lines for biomedical research, including for drug development projects. Such projects often include comparative RNA-sequencing of large panels of human cell lines from individuals affected by certain disorders and healthy controls, or from individuals with different drug response phenotypes. RNA extractions are typically done from growing cell cultures, a process that may take several weeks. However, maintaining large numbers of cell lines in parallel increases the project workload.

Researchers at Tel Aviv University show that extracting RNAs directly from frozen vials of human cell lines stored for over 20 years in a liquid nitrogen freezer yields RNAs with the high purity and integrity parameters that conform to those required for optimal RNA-sequencing and are closely similar to those obtained for RNAs extracted from growing human cell lines.

Comparisons of the RNA integrity numbers (RIN) and RNA purity parameters of the three cohorts as measured by the TapeStation and Qubit systems for human lymphoblastoid cell lines

(a) RIN number: a one-way ANOVA test was applied with significant results of variance between groups (p = .0039), with a following Dunn’s multiple comparisons test showing a significant RIN difference between Cohorts 1 and 3 (p = .002). (b) RNA purity: a one-way ANOVA test was applied with significant results of variance between groups (p = .011), with a following Dunn’s multiple comparisons test showing a significant RIN difference between Cohorts 1 and 2 (p = .02). For both RIN and purity measurements, p < 0.05 were considered significant. ANOVA, analysis of variance