Dataset Information

Robust continuous in vitro culture of the Plasmodium cynomolgi erythrocytic stages.

ABSTRACT: The ability to culture pathogenic organisms substantially enhances the quest for fundamental knowledge and the development of vaccines and drugs. Thus, the elaboration of a protocol for the in vitro cultivation of the erythrocytic stages of Plasmodium falciparum revolutionized research on this important parasite. However, for P. vivax, the most widely distributed and difficult to treat malaria parasite, a strict preference for reticulocytes thwarts efforts to maintain it in vitro. Cultivation of P. cynomolgi, a macaque-infecting species phylogenetically close to P. vivax, was briefly reported in the early 1980s, but not pursued further. Here, we define the conditions under which P. cynomolgi can be adapted to long term in vitro culture to yield parasites that share many of the morphological and phenotypic features of P. vivax. We further validate the potential of this culture system for high-throughput screening to prime and accelerate anti-P. vivax drug discovery efforts.

SUBMITTER: Chua ACY

PROVIDER: S-EPMC6690977 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Publications

Robust continuous in vitro culture of the Plasmodium cynomolgi erythrocytic stages.

Chua Adeline C Y ACY Ong Jessica Jie Ying JJY Malleret Benoit B Suwanarusk Rossarin R Kosaisavee Varakorn V Zeeman Anne-Marie AM Cooper Caitlin A CA Tan Kevin S W KSW Zhang Rou R Tan Bee Huat BH Abas Siti Nurdiana SN Yip Andy A Elliot Anne A Joyner Chester J CJ Cho Jee Sun JS Breyer Kate K Baran Szczepan S Lange Amber A Maher Steven P SP Nosten François F Bodenreider Christophe C Yeung Bryan K S BKS Mazier Dominique D Galinski Mary R MR Dereuddre-Bosquet Nathalie N Le Grand Roger R Kocken Clemens H M CHM Rénia Laurent L Kyle Dennis E DE Diagana Thierry T TT Snounou Georges G Russell Bruce B Bifani Pablo P

Nature communications 20190812 1

The ability to culture pathogenic organisms substantially enhances the quest for fundamental knowledge and the development of vaccines and drugs. Thus, the elaboration of a protocol for the in vitro cultivation of the erythrocytic stages of Plasmodium falciparum revolutionized research on this important parasite. However, for P. vivax, the most widely distributed and difficult to treat malaria parasite, a strict preference for reticulocytes thwarts efforts to maintain it in vitro. Cultivation of ...[more]

PMID: 31406175

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Project description:The Plasmodium falciparum in vitro culture system is critical for genotypic and phenotypic analyses of the parasites. For genotypic analysis, the genomic DNA can be obtained directly from the patient blood sample or from culture adapted parasites whereas for phenotypic analysis, immediate ex vivo or in vitro culture adapted parasites are used. However, parasite biology studies have not investigated whether culture adaptation process affects genotypic and/or phenotypic characteristics of the parasites in short- or long-term cultures. Here, we set out to study the dynamics and stability of parasite genetic and phenotypic profiles as field isolate parasites were adapted in continuous cultures. Parasites collected from three different patients presenting with uncomplicated malaria were adapted and maintained in drug-free continuous cultures. Aliquots from the continuous cultures were collected every 24-48 hours for analyses. Each aliquot was treated as a separate parasite sample. For genetic analysis, microsatellite (MS) typing and single nucleotide polymorphism (SNP) analyses of 23 drug resistance markers were done. The 50% inhibitory concentrations (IC50) for some of the samples were also established for four antimalarial drugs. Samples from each patient (parasite-line) were compared as they were passed through the continuous culture. Data revealed genotypic and phenotypic profiles for the three parasite-lines fluctuated from one generation to the next with no specific pattern or periodicity. With few exceptions, multilocus analysis revealed samples from each parasite-line had high genetic diversity with unique haplotypes. Interestingly, changes in MS and SNP profiles occurred simultaneously. The difference in the IC50s of samples in each parasite-line reached statistical significance. However, phenotypic changes did not correspond or correlate to genotypic changes. Our study revealed parasite genetic and phenotypic characteristics fluctuates in short- and long-term cultures, which indicates parasite genetic information obtained even in short cultures is likely to be different from the natural infection parasites.

Dataset Information

Robust continuous in vitro culture of the Plasmodium cynomolgi erythrocytic stages.

Publications

Robust continuous in vitro culture of the Plasmodium cynomolgi erythrocytic stages.

Similar Datasets

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