Small-scale in vitro culture and purification of Plasmodium berghei for transfection experiment

• Main Authors: Somsak V., Srichairatanakool S., Kamchonwongpaisan S., Yuthavong Y., Uthaipibull C.
• Format: Article
• Language: English
• Published: 2014
• Online Access: http://www.scopus.com/inward/record.url?eid=2-s2.0-79953903955&partnerID=40&md5=9a4d34e211d4149fbc6265ce1365835c; http://cmuir.cmu.ac.th/handle/6653943832/3753
• Institution: Chiang Mai University

The standard protocol for genetic modification of the rodent malaria parasite Plasmodium berghei requires infected blood from one or more laboratory mice, followed by large-scale in vitro parasite culture and purification of mature schizonts. Here, protocols are described for small-scale in vitro culture from 20 μL of mouse tail blood and purification of mature P. berghei schizonts sufficient for a single transfection experiment. All procedures are performed in 1.5-mL microcentrifuge tubes. We confirmed that transgenic parasites could be obtained using schizonts prepared by this protocol. This small-scale protocol provides significant advantages, namely reduction of parasite sample, laboratory consumables and mice for transfection experiments. © 2011 Elsevier B.V. All rights reserved.