Graduation Year

2014

Document Type

Dissertation

Degree

Dr.P.H.

Degree Name

Doctor of Public Health (Dr.PH.)

Degree Granting Department

Global Health

Major Professor

John H. Adams, Ph.D.

Co-Major Professor

Wajeeh M. Saadi, Ph.D.

Committee Member

Dennis E. Kyle, Ph.D.

Committee Member

Thomas R. Unnasch, Ph.D.

Committee Member

Joe D. Cuiffi, Ph.D.

Keywords

hepatocyte, hypnozoite, liver, multiplexed platform, Plasmoidum

Abstract

Malaria is a critical and global public health problem, affecting over 200 million people every year, resulting in over 500,000 deaths. A vaccine is not currently available and only one drug, primaquine, is effective against the dormant stages of Plasmodium vivax. Preclinical assessment of novel therapeutic drugs and vaccines is hampered by the lack of an in vitro liver model for P. falciparum and P. vivax. To provide a stable human hepatocyte-based culture platform for parasite development, we engineered a microfluidic bilayer device capable of both simple and complex culture methods, including perfusion and co-culture, to better understand the requirements of both hepatocytes as host cells and parasite liver stage development. As only the mechanical compaction of the device channels was found necessary for stable hepatocyte culture, and only a validated host cell lot was found necessary for improved parasite development rates, we present an efficient and simple Plasmodium liver model capable of supporting P. vivax dormant forms. Device liver platforms were used to generate kill curves of P. vivax and P. falciparum after three days under primaquine drug pressure. Furthermore, an anti-CSP antibody-based inhibition of development assay with P. falciparum successfully demonstrated antibody-specific liver stage inhibition. Ongoing studies aim to identify the minimal unit of the culture system necessary to be multiplexed in a fully functional and efficient drug and vaccine discovery platform.

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