Optimized in vitro production of Plasmodium vivax ookinetes.
Previous reports have described obtaining mature Plasmodium vivax ookinetes in vitro using blood from infected patients using a simplified, field-based protocol. Here, we report protocols that produce improved P. vivax ookinete yields and morphological development. Optimal conditions included induction of gametogenesis using 10 mM Tris, 170 mM NaCl, 10 mM glucose, 25 mM NaHCO(3), and 100 μM xanthurenic acid for 90 minutes at pH 8.0-8.2, followed by culture in RPMI-1640, 50 mg/mL hypoxanthine, 25 mM HEPES, 29 mM NaHCO(3), 2 mM L-glutamine, and 20% fetal bovine serum at pH 8.4 for 36 hours. Ookinetes were produced in 86% (18/21) of optimized in vitro cultures; yields ranged from 6.5 × 10(4) to 2.8 × 10(6); percent gametocyte conversion ranged from 1.4% to 4.7%. This improved method is suitable for preparation of P. vivax ookinetes in quantities sufficient for biochemical, molecular, and cell biological analysis where basic laboratory facilities are in proximity to patients with vivax malaria.
Division of Infectious Diseases, Department of Medicine, University of California San Diego, La Jolla, California 92093-0741, USA. cmmcclean@gmail.com
Colleen M
CM
McClean
Haydee Guerra
HG
Alvarado
Victor
V
Neyra
Alejandro
A
Llanos-Cuentas
Joseph M
JM
Vinetz
D000818:Animals; D001769:Blood / Q000469:parasitology; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media; D006801:Humans; D016780:Malaria, Vivax; D009866:Oogenesis; D010966:Plasmodium vivax / Q000502:physiology*; D012098:Reproduction / Q000502:physiology
D003470:Culture Media