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Exp Parasitol. 2014 May;140:18-23. doi: 10.1016/j.exppara.2014.03.010. Epub 2014 Mar 13.

Refrigeration provides a simple means to synchronize in vitro cultures of Plasmodium falciparum.

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  • 1Department of Pathogen Biology and Immunology, Kunming Medical University, Yunnan Province 650500, China.
  • 2Department of Pharmacology, Kunming Medical University, Yunnan Province 650500, China.
  • 3Department of Pathogen Biology and Immunology, Kunming Medical University, Yunnan Province 650500, China; Department of Pharmacology, Kunming Medical University, Yunnan Province 650500, China.
  • 4Animal Parasitic Disease Laboratory, Agricultural Research Service, US Department of Agriculture, BARC East Building 1180, Beltsville, MD 20705, USA.
  • 5School of Public Health, Kunming Medical University, Yunnan Province 650500, China.
  • 6Department of Cell Biology and Genetics, Kunming Medical University, Yunnan Province 650500, China.
  • 7Center for Biomedical Engineering Research, Kunming Medical University, Yunnan Province 650500, China.
  • 8Department of Entomology, The Pennsylvania State University, 501 ASI Building, University Park, PA 16802, USA. Electronic address: luc2@psu.edu.
  • 9Department of Pharmacology, Kunming Medical University, Yunnan Province 650500, China. Electronic address: zhaoqingy@yahoo.com.

Abstract

Plasmodium falciparum is usually asynchronous during in vitro culture. Highly synchronized cultures of P. falciparum are routinely used in malaria research. Here, we describe a simple synchronization procedure for P. falciparum asexual erythrocytic culture, which involves storage at 4°C for 8-24 h followed by routine culture. When cultures with 27-60% of ring stage were synchronized using this procedure, 70-93% ring stages were obtained after 48 h of culture and relative growth synchrony remained for at least two erythrocytic cycles. To test the suitability of this procedure for subsequent work, drug sensitivity assays were performed using four laboratory strains and four freshly adapted clinical P. falciparum isolates. Parasites synchronized by sorbitol treatment or refrigeration showed similar dose-response curves and comparable IC50 values to four antimalarial drugs. The refrigeration synchronization method is simple, inexpensive, time-saving, and should be especially useful when large numbers of P. falciparum culture are handled.

Copyright © 2014 Elsevier Inc. All rights reserved.

KEYWORDS:

Drug susceptibility; Plasmodium falciparum; Refrigeration; Synchronization

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