94 Scopus citations


Despite being a frequent cause of severe diarrheal disease in infants and an opportunistic infection in immunocompromised patients, Cryptosporidium research has lagged due to a lack of facile experimental methods. Here, we describe a platform for complete life cycle development and long-term growth of C. parvum in vitro using “air-liquid interface” (ALI) cultures derived from intestinal epithelial stem cells. Transcriptomic profiling revealed that differentiating epithelial cells grown under ALI conditions undergo profound changes in metabolism and development that enable completion of the parasite life cycle in vitro. ALI cultures support parasite expansion > 100-fold and generate viable oocysts that are transmissible in vitro and to mice, causing infection and animal death. Transgenic parasite lines created using CRISPR/Cas9 were used to complete a genetic cross in vitro, demonstrating Mendelian segregation of chromosomes during meiosis. ALI culture provides an accessible model that will enable innovative studies into Cryptosporidium biology and host interactions. Wilke et al. describe an air-liquid interface (ALI) cultivation system that permits the efficient growth of C. parvum in vitro. ALI culture supports life cycle completion, thereby generating oocysts that can propagate in vitro, cause infection in mice, and enable in vitro genetic crosses, thus opening this system for future studies.

Original languageEnglish
Pages (from-to)123-134.e8
JournalCell Host and Microbe
Issue number1
StatePublished - Jul 10 2019


  • Mendelian genetics
  • development
  • host-pathogen interactions
  • meiosis
  • organoids
  • pathway analysis
  • stem cells
  • transcriptomics


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