Culture for genetic manipulation of developmental stages of Schistosoma mansoni.
Genomes of the major human helminth parasites, and indeed many others of agricultural significance, are now the research focus of intensive genome sequencing and annotation. A draft genome sequence of the filarial parasite Brugia malayi was reported in 2007 and draft genomes of two of the human schistosomes, Schistosoma japonicum and S. mansoni reported in 2009. These genome data provide the basis for a comprehensive understanding of the molecular mechanisms involved in schistosome nutrition and metabolism, host-dependent development and maturation, immune evasion and invertebrate evolution. In addition, new potential vaccine candidates and drug targets will likely be predicted. However, testing these predictions is often not straightforward with schistosomes because of the difficulty and expense in maintenance of the developmental cycle. To facilitate this goal, several developmental stages can be maintained in vitro for shorter or longer intervals of time, and these are amenable to manipulation. Our research interests focus on experimental studies of schistosome gene functions, and more recently have focused on development of transgenesis and RNA interference with the longer term aim of heritable gene manipulation. Here we review methods to isolate and culture developmental stages of Schistosoma mansoni, including eggs, sporocysts, schistosomules and adults, in particular as these procedures relate to approaches for gene manipulation. We also discuss recent advances in genetic manipulation of schistosomes including the deployment of square wave electroporation to introduce reporter genes into cultured schistosomes.
Mann
Victoria H
VH
Department of Microbiology, Immunology & Tropical Medicine, George Washington University Medical Center, Washington, DC 20037, USA. mtmvhm@gwumc.edu
Morales
Maria E
ME
Rinaldi
Gabriel
G
Brindley
Paul J
PJ
D000818:Animals; D018929:Cell Culture Techniques; D006801:Humans; D033661:Oocysts / Q000166:cytology* / Q000502:physiology; D010063:Ovum / Q000166:cytology / Q000502:physiology; D012550:Schistosoma mansoni / Q000235:genetics* / Q000254:growth & development* / Q000302:isolation & purification