Continuous culture of Cryptosporidium parvum using hollow fiber technology

Mary Morada, Sangun Lee, Leslie Gunther-Cummins, Louis M. Weiss, Giovanni Widmer, Saul Tzipori, Nigel Yarlett

Research output: Contribution to journalArticle

58 Scopus citations

Abstract

Diarrheal disease is a leading cause of pediatric death in economically low resource countries. Cryptosporidium spp. are the second largest member of this group and the only member for which no treatment exists. One of the handicaps to developing chemotherapy is the lack of a reproducible long-term culture method permitting in vitro drug screening beyond 48 h. We have adapted the well-established hollow fiber technology to provide an environment that mimics the gut by delivering nutrients and oxygen from the basal layer upwards while allowing separate redox and nutrient control of the lumen for parasite development. Using this technique, oocyst production was maintained for >6 months, producing approximately 1 × 108 oocysts ml-1 day-1, compared with 48 h with a yield of 1 × 106 oocysts ml-1 in two-dimensional cultures. Oocysts, after 4 and 20 weeks in culture, produced a chronic infection in a TCR-α-deficient mouse model. In vivo infectivity of oocysts was confirmed using oocysts from a 6 week culture in a dexamethasone immunosuppressed mouse model.

Original languageEnglish (US)
Pages (from-to)21-29
Number of pages9
JournalInternational Journal for Parasitology
Volume46
Issue number1
DOIs
StatePublished - 2016

Keywords

  • Anaerobic
  • Cryptosporidium
  • Hollow-fiber
  • In vitro culture

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

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