Culture of cells from juvenile worms of Schistosoma mansoni.
Tissue disruption methods were developed and serum-free cell culture media formulated for the maintenance in vitro of cells from juvenile worms (day 18 after infection) of Schistosoma mansoni. Cultures maintained viability for up to 6 mo when plated on a feeder layer of irradiated rat liver cells and survived primarily as clusters of small (2.5-4 microns diameter) cells with a high nuclear-to-cytoplasmic ratio and relatively few organelles identified by electron microscopy. Cultures synthesized a protein profile similar to that of intact worms, and the cell clusters maintained a time- and concentration-dependent contractile response to serotonin. Cells synthesizing DNA were detected by precursor incorporation and flow cytometry in cultures initially and also after several weeks in vitro, although the percentage of cells synthesizing DNA decreased with time. Efforts to identify peptide growth factor-responsive tyrosine phosphorylation were negative, and the overall amount of S. mansoni phosphotyrosine-containing proteins identified by western blot with anti-phosphotyrosine monoclonal antibody was much less than that found in a peptide growth factor-responsive mouse cell line.
Hobbs
D J
DJ
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-6503.
Fryer
S E
SE
Duimstra
J R
JR
Hedstrom
O R
OR
Brodie
A E
AE
Collodi
P A
PA
Menino
J S
JS
Bayne
C J
CJ
Barnes
D W
DW
D000818:Animals; D015153:Blotting, Western; D002448:Cell Adhesion; D002460:Cell Line; D016895:Culture Media, Serum-Free; D004247:DNA / Q000096:biosynthesis; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005434:Flow Cytometry; D015801:Helminth Proteins / Q000096:biosynthesis; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D012550:Schistosoma mansoni / Q000166:cytology / Q000254:growth & development* / Q000648:ultrastructure
D016895:Culture Media, Serum-Free; D015801:Helminth Proteins; D004815:Epidermal Growth Factor; D004247:DNA