The MTT Assay: A Method for Error Minimization and Interpretation in Measuring Cytotoxicity and Estimating Cell Viability.

The MTT assay is extensively used, most often to infer a measure of cytotoxicity of treatments to cells. As with any assay though, there are a number of limitations. The method described here is designed with consideration of how the MTT assay fundamentally works to account for, or at least identify, confounding factors in measurements. It also provides a decision-making framework to best interpret and complement the MTT assay to apply it as either a measure of metabolic activity or cell viability.

Characteristic features of porcine endogenous retroviruses in Vietnamese native pigs.

We aimed to clarify the genomic characteristics of porcine endogenous retroviruses (PERVs) in Vietnamese native pig (VnP) breeds. First, we investigated genetic polymorphisms in ß- and γ-like PERVs, and we then measured the copy numbers of infectious γ-like PERVs (PERV-A, B, and C). We purified genomic DNA from 15 VnP breeds from 12 regions all over the country and three Western pig breeds as controls, and investigated genetic polymorphisms in all known PERVs, including the beta (ß)1-4 and gamma (γ)1-5 groups. PERVs of ß1, ß2, ß3, and γ4 were highly polymorphic with VnP-specific haplotypes. We did not identify genetic polymorphisms in ß4, γ1, or γ2 PERVs. We then applied a real-time polymerase chain reaction-based method to estimate copy numbers of the gag, pol, and env genes of γ1 PERVs (defined as A, B, and C). VnP breeds showed significantly lower copy number of the PERV genes compared with the Western pig breeds (on average, 16.2 and 35.7 copies, respectively, p < .05). Two VnP breeds showed significantly higher copy number compared with the other VnPs (p < .05). Our results elucidated that VnPs have specific haplotypes and a low copy number of PERV genes.