Cytometric analysis, genetic manipulation and antibiotic selection of the snail embryonic cell line Bge from Biomphalaria glabrata, the intermediate host of Schistosoma mansoni

Gabriel Rinaldi, Hongbin Yan, Rafael Nacif-Pimenta, Pitchaya Matchimakul, Joanna Bridger, Victoria H. Mann, Michael J. Smout, Paul J. Brindley, Matty Knight*

*Awdur cyfatebol y gwaith hwn

Allbwn ymchwil: Cyfraniad at gyfnodolynErthygladolygiad gan gymheiriaid

12 Dyfyniadau (Scopus)

Crynodeb

The invertebrate cell line, Bge, from embryos of the snail Biomphalaria glabrata, remains to date the only established cell line from any species of the Phylum Mollusca. Since its establishment in 1976 by Eder Hansen, few studies have focused on profiling its cytometrics, growth characteristics or sensitivity to xenobiotics. Bge cells are reputed to be challenging to propagate and maintain. Therefore, even though this cell line is a noteworthy resource, it has not been studied widely. With growing interest in functional genomics, including genetic transformation, to elucidate molecular aspects of the snail intermediate hosts responsible for transmission of schistosomiasis, and aiming to enhance the convenience of maintenance of this molluscan cell line, we deployed the xCELLigene real time approach to study Bge cells. Doubling times for three isolates of Bge, termed CB, SL and UK, were longer than for mammalian cell lines - longer than 40 h in complete Bge medium supplemented with 7% fetal bovine serum at 25. °C, ranging from ~42 h to ~157 h when 40,000 cells were seeded. To assess the potential of the cells for genetic transformation, antibiotic selection was explored. Bge cells were sensitive to the aminonucleoside antibiotic puromycin (from Streptomyces alboniger) from 5. μg/ml to 200. ng/ml, displaying a half maximal inhibitory concentration (IC50) of ~1.91 μg/ml. Sensitivity to puromycin, and a relatively quick kill time (<48 h in 5. μg/ml) facilitated use of this antibiotic, together with the cognate resistance gene (puromycin N-acetyl-transferase) for selection of Bge cells transformed with the PAC gene (puroR). Bge cells transfected with a plasmid encoding puroR were partially rescued when cultured in the presence of 5. μg/ml of puromycin. These findings pave the way for the development of functional genomic tools applied to the host-parasite interaction during schistosomiasis and neglected tropical trematodiases at large.

Iaith wreiddiolSaesneg
Tudalennau (o-i)527-535
Nifer y tudalennau9
CyfnodolynInternational Journal for Parasitology
Cyfrol45
Rhif cyhoeddi8
Dynodwyr Gwrthrych Digidol (DOIs)
StatwsCyhoeddwyd - 01 Gorff 2015
Cyhoeddwyd yn allanolIe

Ôl bys

Gweld gwybodaeth am bynciau ymchwil 'Cytometric analysis, genetic manipulation and antibiotic selection of the snail embryonic cell line Bge from Biomphalaria glabrata, the intermediate host of Schistosoma mansoni'. Gyda’i gilydd, maen nhw’n ffurfio ôl bys unigryw.

Dyfynnu hyn