TY - JOUR
T1 - Evaluating the use of heparin for synchronization of in vitro culture of Plasmodium falciparum
AU - Kobayashi, Kyousuke
AU - Kato, Kentaro
N1 - Funding Information:
This study was supported by a JSPS Research Fellowship for Young Scientists ( 11J08985 ) and by the Bio-oriented Technology Research Advancement Institution (BRAIN).
PY - 2016/10
Y1 - 2016/10
N2 - The malaria parasite Plasmodium falciparum infects human erythrocytes and reproduces asexually through an intraerythrocytic developmental cycle. In vitro culture of P. falciparum allows investigation of the parasite's blood-stage development, which spans approximately 48 h from the time of invasion to the lysis of mature schizonts to release merozoites. To focus on a specific step in the developmental cycle, synchronization techniques are utilized. D-Sorbitol treatment and the Percoll-sorbitol method have been used; however, these techniques have limitations in terms of the degree of synchronization achieved, the amount of synchronized parasite acquired, convenience, reproducibility, and cost. Here, we evaluated an existing synchronization method involving heparin. Heparin reversibly inhibits erythrocyte invasion by P. falciparum merozoites. We confirm that parasite cultures can be inexpensively, reproducibly, and tightly synchronized by combining a sorbitol step to limit cultures to the ring stages and by adding and removing heparin to manipulate the window during which merozoites can invade erythrocytes.
AB - The malaria parasite Plasmodium falciparum infects human erythrocytes and reproduces asexually through an intraerythrocytic developmental cycle. In vitro culture of P. falciparum allows investigation of the parasite's blood-stage development, which spans approximately 48 h from the time of invasion to the lysis of mature schizonts to release merozoites. To focus on a specific step in the developmental cycle, synchronization techniques are utilized. D-Sorbitol treatment and the Percoll-sorbitol method have been used; however, these techniques have limitations in terms of the degree of synchronization achieved, the amount of synchronized parasite acquired, convenience, reproducibility, and cost. Here, we evaluated an existing synchronization method involving heparin. Heparin reversibly inhibits erythrocyte invasion by P. falciparum merozoites. We confirm that parasite cultures can be inexpensively, reproducibly, and tightly synchronized by combining a sorbitol step to limit cultures to the ring stages and by adding and removing heparin to manipulate the window during which merozoites can invade erythrocytes.
KW - Heparin
KW - In vitro culture
KW - Plasmodium falciparum
KW - Synchronization
UR - http://www.scopus.com/inward/record.url?scp=84995422439&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84995422439&partnerID=8YFLogxK
U2 - 10.1016/j.parint.2016.09.002
DO - 10.1016/j.parint.2016.09.002
M3 - Article
AN - SCOPUS:84995422439
VL - 65
SP - 549
EP - 551
JO - Parasitology International
JF - Parasitology International
SN - 1383-5769
IS - 5
ER -