Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax

© 2015 Roobsoong et al. Background: The study of the biology, transmission and pathogenesis of Plasmodium vivax is hindered due to the lack of a robustly propagating, continuous culture of this parasite. The current culture system for P. vivax parasites still suffered from consistency and difficulti...

Full description

Saved in:
Bibliographic Details
Main Authors: Wanlapa Roobsoong, Chayada S. Tharinjaroen, Nattawan Rachaphaew, Porpimon Chobson, Louis Schofield, Liwang Cui, John H. Adams, Jetsumon Sattabongkot
Format: Journal
Published: 2018
Subjects:
Online Access:https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84938598830&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/44255
Tags: Add Tag
No Tags, Be the first to tag this record!
Institution: Chiang Mai University
id th-cmuir.6653943832-44255
record_format dspace
spelling th-cmuir.6653943832-442552018-04-25T07:47:27Z Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax Wanlapa Roobsoong Chayada S. Tharinjaroen Nattawan Rachaphaew Porpimon Chobson Louis Schofield Liwang Cui John H. Adams Jetsumon Sattabongkot Agricultural and Biological Sciences © 2015 Roobsoong et al. Background: The study of the biology, transmission and pathogenesis of Plasmodium vivax is hindered due to the lack of a robustly propagating, continuous culture of this parasite. The current culture system for P. vivax parasites still suffered from consistency and difficulties in long-term maintenance of parasites in culture and for providing sufficient biological materials for studying parasite biology. Therefore, further improvement of culture conditions for P. vivax is needed. Methods: Clinical samples were collected from patients diagnosed with P. vivax in western Thailand. Leukocyte-depleted P. vivax infected blood samples were cultured in a modified McCoy's 5A medium at 5% haematocrit under hypoxic condition (5% O < inf > 2 < /inf > , 5% CO < inf > 2 < /inf > , and 90% N < inf > 2 < /inf > ). Reticulocytes purified from adult peripheral blood were added daily to maintain 4% reticulocytes. Parasites were detected by microscopic examination of Giemsa-stained smears and molecular methods. Results: The effects of culture variables were first analysed in order to improve the culture conditions for P. vivax. Through analysis of the sources of host reticulocytes and nutrients of culture medium, the culture conditions better supporting in vitro growth and maturation of the parasites were identified. Using this system, three of 30 isolates could be maintained in vitro for over 26 months albeit parasite density is low. Conclusions: Based on the analysis of different culture variables, an improved and feasible protocol for continuous culture of P. vivax was developed. 2018-01-24T04:39:58Z 2018-01-24T04:39:58Z 2015-08-05 Journal 14752875 2-s2.0-84938598830 10.1186/s12936-015-0815-z https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84938598830&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/44255
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Agricultural and Biological Sciences
spellingShingle Agricultural and Biological Sciences
Wanlapa Roobsoong
Chayada S. Tharinjaroen
Nattawan Rachaphaew
Porpimon Chobson
Louis Schofield
Liwang Cui
John H. Adams
Jetsumon Sattabongkot
Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax
description © 2015 Roobsoong et al. Background: The study of the biology, transmission and pathogenesis of Plasmodium vivax is hindered due to the lack of a robustly propagating, continuous culture of this parasite. The current culture system for P. vivax parasites still suffered from consistency and difficulties in long-term maintenance of parasites in culture and for providing sufficient biological materials for studying parasite biology. Therefore, further improvement of culture conditions for P. vivax is needed. Methods: Clinical samples were collected from patients diagnosed with P. vivax in western Thailand. Leukocyte-depleted P. vivax infected blood samples were cultured in a modified McCoy's 5A medium at 5% haematocrit under hypoxic condition (5% O < inf > 2 < /inf > , 5% CO < inf > 2 < /inf > , and 90% N < inf > 2 < /inf > ). Reticulocytes purified from adult peripheral blood were added daily to maintain 4% reticulocytes. Parasites were detected by microscopic examination of Giemsa-stained smears and molecular methods. Results: The effects of culture variables were first analysed in order to improve the culture conditions for P. vivax. Through analysis of the sources of host reticulocytes and nutrients of culture medium, the culture conditions better supporting in vitro growth and maturation of the parasites were identified. Using this system, three of 30 isolates could be maintained in vitro for over 26 months albeit parasite density is low. Conclusions: Based on the analysis of different culture variables, an improved and feasible protocol for continuous culture of P. vivax was developed.
format Journal
author Wanlapa Roobsoong
Chayada S. Tharinjaroen
Nattawan Rachaphaew
Porpimon Chobson
Louis Schofield
Liwang Cui
John H. Adams
Jetsumon Sattabongkot
author_facet Wanlapa Roobsoong
Chayada S. Tharinjaroen
Nattawan Rachaphaew
Porpimon Chobson
Louis Schofield
Liwang Cui
John H. Adams
Jetsumon Sattabongkot
author_sort Wanlapa Roobsoong
title Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax
title_short Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax
title_full Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax
title_fullStr Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax
title_full_unstemmed Improvement of culture conditions for long-term in vitro culture of Plasmodium vivax
title_sort improvement of culture conditions for long-term in vitro culture of plasmodium vivax
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84938598830&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/44255
_version_ 1681422525651746816