Application of ribonucleoside vanadyl complex (RVC) for developing a multifunctional tissue preservative solution

Abstract

The quality of biological samples greatly affects the accuracy of scientific results. However, RNA in cryopreserved tissues gradually degrades during storage, leading to errors in the results of subsequent experiments. A suitable sample preservative solution can prolong storage and enhance the research value of samples. Here, we developed a sample preservative solution using the properties of the ribonucleoside vanadyl complex (RVC) and compared its effects on RNA and DNA quality, protein activity, and tissue morphology with the commercially available and widely used RNAlater® Stabilization Solution. The results showed that both the RVC-based preservative solution and RNAlater can effectively delay RNA degradation in tissue samples stored at 4°C or −80°C compared with samples stored without any preservative solution. In contrast to RNAlater, the RVC-based preservative solution did not result in damage to the tissue morphology or a loss of protein activity. Additionally, the RVC-based preservative solution did not affect the RNA and genomic DNA contents of the tissue samples or the results of subsequent experimental analyses. An RVC-based reagent can be used as a multifunctional yet relatively inexpensive tissue preservative solution to provide a comprehensive and cost-effective method for preserving samples for tissue banks.

Robust single nucleus RNA sequencing reveals depot-specific cell population dynamics in adipose tissue remodeling during obesity

“…Further analysis of nucleus morphology at a higher resolution revealed a smoother membrane surface and larger nucleus size when using the optimized protocol, as compared to the standard protocol ( Figure 1F ). Consistent with the preservative function of VRC proven in maintaining tissue morphology 26 , these findings show that our nucleus isolation protocol effectively maintains nucleus structure without shrinkage. Given that current snRNAseq methods require prompt sample processing due to the rapid degradation of RNA in isolated nuclei, we further tested the temporal stability of RNA in nuclei isolated using our optimized protocol.…”

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH

“…However, adjusting fixation time and temperature is necessary for your specific sample to avoid over-or under-fixing the sample 26 . VRC is useful because it reduces RNA degradation and is helpful especially for low abundant RNA 27 . There are two options that we have had success with for the permeabilization step.…”

Genomics and metatranscriptomics of biogeochemical cycling and degradation of lignin-derived aromatic compounds in thermal swamp sediment

“…RNases from thermophilic organisms are highly resistant to inhibition or denaturation. To recover high-quality RNA from thermophilic communities, 100 µl of 200 mM ribonucleoside vanadyl complex (RVC) (Sigma Aldrich) was added to the extraction buffer [ 37 , 38 ]. RVC was essential for recovery of RNA from these samples (Supplementary Fig.…”