Immortalization of shrimp lymphoid cells by hybridizing with the continuous cell line Sf9 leading to the development of ‘PmLyO-Sf9 ’

• Published in: Fish & shellfish immunology Vol. 113; pp. 196 - 207
• Main Authors: Anoop, B.S., Puthumana, Jayesh, Vazhappilly, Cijo George, Kombiyil, Salini, Philip, Rosamma, Abdulaziz, Anas, Bright Singh, Isaac Sarojini
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01-06-2021
• Subjects: Cell fusion and TEM; Hybrid cell line; Sf9 cells; Shrimp cell culture; Hybrid cell line; Shrimp cell culture; Cell fusion and TEM; Sf9 cells
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2021.03.023

Shrimp progressively gets more attention among marine invertebrates from researchers all over the world due to it being a healthy food as well as having economic importance. There were a lot of attempts to develop a continuous cell line from shrimp but none successful. In this context a novel hybrid cell line named ‘PmLyO-Sf9’ could be developed by fusing shrimp lymphoid organ cells with Sf9 cells after to metabolic blocking of Sf9 cells using puromycin and actinomycin D and effecting the fusion by way of PEG application. The cells are maintained and multiplied in a mixture of SCCM and TNM-FH having osmolality 550 mOsm kg−1 and pH 6.8. Transmission electron microscopy of the hybrid cells revealed the presence of two nuclei during the initial stages and a single nucleus subsequently. The cell line is with shrimp and Sf9 genomic components and shrimp specific protein and is susceptible to WSSV. Shrimp elongation factor, Sf9 beta-actin, shrimp STAT and peroxinectin could be expresses through RT-PCR in the cell line. This is the first successful report of a hybrid cell line with shrimp genomic components and envisaged to be recognized a model system for multitudes of biomedical research in vitro. The cell line is in the National Cell Line Repository of ICAR – National Bureaue of Fish Genetic Resources, Lucknow, India.
•A novel hybrid cell line ‘PmLyO-Sf9’ developed by fusing shrimp lymphoid organ cells with Sf9 cells.•Morphologically, hybrid cells are bi- and tri-polar with tail like extension.•Electron microscopy analysis revealed two distinct nuclei in early stages.•Shrimp and Sf9 genomic components were separately amplified.•Shrimp immune genes peroxinectin and STAT expression confirmed in hybrid cell line.