Comparative evaluation of culture media for improved yield and viability of primary testicular cell lines from ovine (lamb) tissue

Abstract

Primary cell culture is the ex vivo culture of cells freshly obtained from a multicellular organism, as opposed to the culture of immortalized cell lines. Primary cell cultures derived from testicular tissue serve as vital in vitro model for studying viral isolation and dynamics in wide range of viruses. Establishing stable primary testicular cell lines from lambs is often challenging due to variability in growth conditions, contamination risks, and lack of optimized culture media protocols. Selection of appropriate basal media and supplementation strategies plays a critical role in improving cell viability, proliferation, and maintenance of cellular morphology. However, there is limited comparative data on the efficiency of different media formulations for lamb testicular cell culture. LT cells are suitable for most of pox viruses, especially Lumpy Skin Disease virus. It may be noted that adapting a virus to primary cells yields more physiologically relevant results and more accurate model of in vivo infection for sequencing but researchers and vaccinologists are more conveniently dependent on the continuous cell lines for baseline studies and vaccine development. Hence, this study was designed to identify an optimized culture protocol for lamb testicular primary cell line preparation in two of the common cell culture media in varying serum concentration. Two commonly used laboratory media, DMEM and MEM with Earle’s salt were used for the study. It was observed that DMEM supplemented with 10 % FBS could be an efficient and suitable medium for the propagation of the LT primary culture. The standardized protocol may help in the establishment of LT primary culture in the laboratory, understanding viral dynamics in the host, and evolutionary studies.