Transformation of primary cultures of shrimp (Penaeus stylirostris) lymphoid (Oka) organ with Simian virus-40 (T) antigen.
Primary cultures of lymphoid (Oka) organ from Penaeus stylirostris were transformed with naked or Lipofectin-mediated pSV-3 neo, a shuttle vector containing the tumor (T) antigen gene from Simian virus-40. The transformed cells, OKTr-1 and OKTr-23, exhibited the following characteristics: rounded morphology forming grapelike aggregates, loosely adhesive, increased growth rate in Medium-199, resistance to G-418 (a neomycin analog marker in the shuttle vector), cloning efficiencies of 68.7% and 36.7% in soft agarose, respectively, and stability in liquid nitrogen storage. Immunofluorescence staining (IFA) of the transformed cells using a monoclonal antibody against SV-40 tumor antigen showed positive results. In contrast, primary cell cultures exhibited fibroblast-like morphology and formed a tight, adhesive monolayer on the surface of the culture vessel. They were sensitive to G-418, and showed negative results with IFA. To date, OKTr-1 and OKTr-23 have undergone 44 and 18 passages, respectively. Primary cultures of the lymphoid organ have not been successfully passaged beyond the primary stage.
Tapay
L M
LM
Department of Microbiology, University of Hawaii at Manoa, Honolulu 96822, USA.
Lu
Y
Y
Brock
J A
JA
Nadala
E C
EC
Loh
P C
PC
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis / Q000235:genetics*; D002461:Cell Line, Transformed / Q000166:cytology* / Q000502:physiology; D002472:Cell Transformation, Viral; D008221:Lymphoid Tissue / Q000166:cytology; D033561:Penaeidae / Q000166:cytology*; D010957:Plasmids; D000644:Quaternary Ammonium Compounds; D013539:Simian virus 40; D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D000644:Quaternary Ammonium Compounds; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium