1921
Volume 83, Issue 6
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Abstract.

Previous reports have described obtaining mature ookinetes using blood from infected patients using a simplified, field-based protocol. Here, we report protocols that produce improved ookinete yields and morphological development. Optimal conditions included induction of gametogenesis using 10 mM Tris, 170 mM NaCl, 10 mM glucose, 25 mM NaHCO, 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, 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 cultures; yields ranged from 6.5 × 10 to 2.8 × 10; percent gametocyte conversion ranged from 1.4% to 4.7%. This improved method is suitable for preparation of ookinetes in quantities sufficient for biochemical, molecular, and cell biological analysis where basic laboratory facilities are in proximity to patients with vivax malaria.

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2010-12-06
2017-11-20
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  • Received : 04 Apr 2010
  • Accepted : 17 Aug 2010

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