期刊
NATURE PROTOCOLS
卷 4, 期 12, 页码 1899-1915出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2009.198
关键词
-
资金
- Spanish Ministry of Education and Science [BIO2006-12355, BIO2007-67885]
- UCM (Comunidad de Madrid) [920267]
- The Pasteur Institute of Iran
- AECI-Spanish Ministry of Foreign Affairs
- Universidad de Cartagena (CO)
- Alban Programme of High Level Scholarships for Latin America
- European Union [E06D101036CO, E07D400516CO]
- Spanish Ministry of Education and Science
This protocol describes a method for preparing cultures of Plasmodium falciparum synchronized at any intraerythrocytic stage. Using this method, around 60% parasitized cells may be obtained. On the basis of Trager and Jensen's original continuous culture method, our approach relies on the use of fresh human blood not older than 2 weeks, a low hematocrit between 0.8 and 1.5%, a starting frozen inoculum of 10% ring-stage parasitemia, human serum replaced with AlbuMAX I and alternating sorbitol and Percoll synchronization methods to shorten the cycle window to 4-6 h and reduce sorbitol toxicity. From our synchronized high parasite density cultures, 3-5 ml of infected red blood cells can be obtained in 1 week, corresponding to 1.2 mg of total parasite protein per ml of harvested culture. On the basis of the variables parasitemia and packed cell volume, we provide an equation to accurately calculate the amount of complete medium required every 24 h corrected for the cycle stage and capacity of the culture flask. Ten days suffice to complete the protocol from a frozen stock of parasites.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据