PRIMARY CELL CULTURE OF AEDES ALBOPICTUS MIDGUT CELLS: A PROSPECTIVE MODEL FOR IN VITRO STUDY OF ARBOVIRUSES
Â Objective: Midgut cells play a key role in the propagation of mosquito borne Arboviruses. The existing mosquito cell lines for studying viral pathogenesis are derived either from larvae or from eggs since there is no cell line available from the mosquito midgut. Therefore, to delineate the in situ viral interaction which naturally occurs within the mosquito midgut and represent cellular pathogenesis in human beings, the present work was aimed to develop a primary cell line from the midgut cells of Aedes albopictus.
Methods: The midgut cells of A. albopictus were collected, cultured and incubated at 28Â°C to study the growth after every 24 hrs for 7 days.
Result: The primary cell culture showed an increasing growth pattern of columnar cells up to 48 hrs followed by decrease in cell population afterward. However, the number of stem cells increased significantly throughout the study period, and their population outnumbered the columnar cells after 72 hrs. There was no significant change of goblet cells and regenerating cells which were scanty in number throughout the experiment.
Conclusion: The present method will help to develop the individual cell lines from mosquito midgut and study the host pathogen interaction in arboviral diseases in future.
2. Durbin AP, Mayer SV, Rossi SL, Amaya LI, Ramos CJ, Eong OE, et al. Emergence potential of sylvatic dengue virus Type 4 in the urban transmission cycle is restrained by vaccination and homotypic immunity. J Virol 2013;439(1):34-41.
3. Petersen LR, Marfin AA. Shifting epidemiology of Flaviviridae. J Travel Med 2005;12 Suppl 1:S3-11.
4. Franz AW, Kantor AM, Passarelli AL, Clem RJ. Tissue barriers to arbovirus infection in mosquitoes. Viruses 2015;7(7):3741-67.
5. Walker T, Jeffries CL, Mansfield KL, Johnson N. Mosquito cell lines: History, isolation, availability and application to assess the threat of arboviral transmission in the United Kingdom. Parasit Vectors 2014;7:382.
6. Service M. Medical Entomology for Students. 4th ed. New York: Cambridge University Press; 2008. p. 53-80.
7. Das M, Das MK, Dutta P. Genetic characterization and molecular phylogeny of Aedes albopictus (Skuse) species from Sonitpur district of Assam, India based on COI and ITS1 genes. J Vector Borne Dis 2016;53(3):240-7.
8. Sinha M, Roy E, Das S, Sarkar DB, Nayak D, Khurana A, et al. An observation on direct changes in Aedes albopictus midgut cells by Rhus tox 6C in relation to dengue virus infection. Indian J Res Homoeopath 2016;10:258-65.
9. Hamer GL, Anderson TK, Berry GE, Makohon-Moore AP, Crafton JC, Brawn JD, et al. Prevalence of filarioid nematodes and trypanosomes in American robins and house sparrows, Chicago USA. Int J Parasitol Parasites Wildl 2012;2:42-9.
10. Sadrud-Din S, Loeb M, Hakim R. In vitro differentiation of isolated stem cells from the midgut of Manduca sexta larvae. J Exp Biol 1996;199:319-25.
11. Brackney DE, Scott JC, Sagawa F, Woodward JE, Miller NA, Schilkey FD, et al. C6/36 Aedes albopictus cells have a dysfunctional antiviral RNA interference response. PLoS Negl Trop Dis 2010;4(10):e856.
12. Zieler H, Garon CF, Fischer ER, Shahabuddin M. A tubular network associated with the brush-border surface of the Aedes aegypti midgut: Implications for pathogen transmission by mosquitoes. J Exp Biol 2000;203:1599-611.
13. Fischer ER, Hansen BT, Nair V, Hoyt FH, Dorward DW. Scanning electron microscopy. Curr Protoc Microbiol 2012;Chapter 2:Unit 2B.2.
14. Billingsley PF. Vector-parasite interactions for vaccine development. Int J Parasitol 1994;24:53-8.
15. Shahabuddin M, Cociancich S, Zieler H. The search for novel malaria transmission-blocking targets in the mosquito midgut. Parasitol Today 1998;14(12):493-7.
16. Hakim RS, Caccia S, Loeb M, Smagghe G. Primary culture of insect midgut cells. In Vitro Cell Dev Biol Anim 2009;45(3-4):106-10.
17. Hakim RS, Baldwin KM, Loeb M. The role of stem cells in midgut growth and regeneration. In Vitro Cell Dev Biol Anim 2001;37(6):338-42.
18. Sushama MA, Kumaresan R. Study on in vitro cytotoxicity of papain against liver cancer cell line Hep G2. Int J Pharm Pharm Sci 2014;6(9):160-1.
19. Aishwarya V, Abdulla SS, Dheeba B, Renuka R. In vitro antioxidant and anticancer activity of Cardiospermum halicabum L against EAC cell line. Int J Pharm Pharm Sci 2014;6(8):263-8.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.