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Neuroscience JoVE (Open Access) Citable · DOI

Selection of Plasmodium falciparum Parasites for Cytoadhesion to Human Brain Endothelial Cells

DOI: 10.3791/3122-v
What you'll learn
  • ✓ Culture Plasmodium falciparum parasites and human brain endothelial cells in vitro
  • ✓ Select P. falciparum for cytoadhesion phenotype using endothelial cell binding
  • ✓ Quantify increased parasite binding to endothelial cells after selection
Protocol

An in vitro model for cerebral malaria sequestration is described1. Plasmodium falciparum infected red blood cells are selected for binding to immortalized human brain microvascular endothelial cells. The selected parasites show a distinct phenotype. The selection process can be applied using various P. falciparum strains and endothelial cell lines.

Difficulty
advanced
Total time
~2–3 weeks (iterative selection cycles with continuous culture maintenance)
Model organism
Plasmodium falciparum, human brain microvascular endothelial cells (HBEC-5i)
Biosafety
BSL-2

Steps

1
Prepare and maintain Plasmodium falciparum culture

Culture P. falciparum parasites using standard protocols with routine media changes and maintenance to ensure viable parasitized red blood cells for selection.

▶ 01:14
2
Culture endothelial cells and prepare for selection

Maintain immortalized human brain microvascular endothelial cells (HBEC-5i) in culture and prepare monolayers as substrate for parasite binding experiments.

▶ 02:54
3
Select parasites for cytoadhesion to endothelial cells

Incubate P. falciparum-infected red blood cells with HBEC-5i monolayers to enrich for parasites with high binding affinity, then recover bound parasites through washing and culture.

▶ 04:00
4
Quantify increased binding after selection rounds

Measure and compare binding efficiency of selected parasites to HBEC-5i cells across selection iterations to confirm phenotypic enrichment.

▶ 07:36
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