# Generation and application of immortalized sheep fetal fibroblast cell line

## TL;DR

Scientists created long-lasting sheep cells that can be used to study a virus and develop vaccines.

## Contribution

A new immortalized sheep fetal fibroblast cell line was developed for ORFV research.

## Key findings

- Immortalized cells maintain the same biological functions as primary cells.
- The cells show resistance to apoptosis and enhanced proliferation.
- They are susceptible to ORFV vaccines and suitable for in vitro studies.

## Abstract

Primary sheep fetal fibroblasts (SFFCs) have emerged as a valuable resource for investigating the molecular and pathogenic mechanisms of orf viruses (ORFV). However, their utilization is considerably restricted due to the exorbitant expenses associated with their isolation and culture, their abbreviated lifespan, and the laborious procedure.

In our investigation, the primary SFFCs were obtained and immortalized by introducing a lentiviral recombinant plasmid containing the large T antigen from simian virus 40 (SV40). The expression of fibronectin and vimentin proteins, activity of SV40 large T antigen, cell proliferation assays, and analysis of programmed cell death revealed that the immortalized large T antigen SFFCs (TSFFCs) maintained the same physiological characteristics and biological functions as the primary SFFCs. Moreover, TSFFCs demonstrated robust resistance to apoptosis, extended lifespan, and enhanced proliferative activity compared to primary SFFCs. Notably, the primary SFFCs did not undergo in vitro transformation or exhibit any indications of malignancy in nude mice. Furthermore, the immortalized TSFFCs displayed live ORFV vaccine susceptibility.

Immortalized TSFFCs present valuable in vitro models for exploring the characteristics of ORFV using various techniques. This indicates their potential for secure utilization in future studies involving virus isolation, vaccine development, and drug screening.

The online version contains supplementary material available at 10.1186/s12917-024-04054-3.