Establishment and partial characterisation of a new cell line derived from adult tissues of the tsetse fly Glossina morsitans morsitans

• Published in: Parasites & vectors Vol. 17; no. 1; p. 231
• Main Authors: Bell-Sakyi, Lesley, Haines, Lee R., Petrucci, Giovanni, Beliavskaia, Alexandra, Hartley, Catherine, Khoo, Jing Jing, Makepeace, Benjamin L., Abd-Alla, Adly M. M., Darby, Alistair C.
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 17.05.2024; BioMed Central; BMC
• Subjects: adults; agricultural development; Analysis; Animals; Cell Line; cell lines; Cells; chromosome number; Female; females; fetal bovine serum; genes; Genetic aspects; Glossina morsitans morsitans; haploidy; human health; hypertrophy; Identification and classification; Iflavirus; iflavirus, negevirus; insect; Insect Vectors - virology; insects; Karyotyping; phosphates; RNA; RNA, Ribosomal, 16S - genetics; salivary glands; species; Sub-Saharan Africa; symbionts; Trypanosoma; tsetse; Tsetse Flies - parasitology; Tsetse-flies; viruses; Wolbachia; United States; Sub-Saharan Africa; iflavirus, negevirus; insect; Wolbachia; cell line; tsetse; Glossina morsitans morsitans
• ISSN: 1756-3305; 1756-3305
• DOI: 10.1186/s13071-024-06310-9

Insect cell lines play a vital role in many aspects of research on disease vectors and agricultural pests. The tsetse fly Glossina morsitans morsitans is an important vector of salivarian trypanosomes in sub-Saharan Africa and, as such, is a major constraint on human health and agricultural development in the region. Here, we report establishment and partial characterisation of a cell line, GMA/LULS61, derived from tissues of adult female G. m. morsitans. GMA/LULS61 cells, grown at 28 °C in L-15 (Leibovitz) medium supplemented with foetal bovine serum and tryptose phosphate broth, have been taken through 23 passages to date and can be split 1:1 at 2-week intervals. Karyotyping at passage 17 revealed a predominantly haploid chromosome complement. Species origin and absence of contaminating bacteria were confirmed by PCR amplification and sequencing of fragments of the COI gene and pan-bacterial 16S rRNA gene respectively. However, PCR screening of RNA extracted from GMA/LULS61 cells confirmed presence of the recently described Glossina morsitans morsitans iflavirus and Glossina morsitans morsitans negevirus, but absence of Glossina pallipides salivary gland hypertrophy virus. GMA/LULS61 cells supported infection and growth of 6/7 different insect-derived strains of the intracellular bacterial symbiont Wolbachia. The GMA/LULS61 cell line has potential for application in a variety of studies investigating the biology of G. m. morsitans and its associated pathogenic and symbiotic microorganisms.