In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers

Georgia Wilke, Yi Wang, Soumya Ravindran, Thaddeus Stappenbeck, William Harold Witola, L. David Sibley

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Cryptosporidium parvum has a complex life cycle consisting of asexual and sexual phases that culminate in oocyst formation in vivo. The most widely used cell culture platforms to study C. parvum only support a few days of growth and do not allow the parasite to proceed past the sexual stages to complete oocyst formation. Additionally, these cell culture platforms are mostly adenocarcinoma cell lines, which do not adequately model the parasite’s natural environment in the small intestine. We describe here a method to create primary intestinal epithelial cell monolayers that support long-term C. parvum growth. Monolayers were derived from mouse intestinal stem cells grown as spheroids and plated onto transwells, allowing for separate apical and basolateral compartments. In the apical chamber, the cell growth medium was removed to create an “air–liquid interface” that enhanced host cell differentiation and supported long-term C. parvum growth. The use of primary intestinal cells to grow C. parvum in vitro will be a valuable tool for studying host–parasite interactions using a convenient in vitro model that more closely resembles the natural niche in the intestine.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages351-372
Number of pages22
DOIs
StatePublished - Jan 1 2020

Publication series

NameMethods in Molecular Biology
Volume2052
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Fingerprint

Cryptosporidium parvum
Stem Cells
Oocysts
Growth
Parasites
Cell Culture Techniques
Life Cycle Stages
Small Intestine
Intestines
Cell Differentiation
Adenocarcinoma
Epithelial Cells
In Vitro Techniques
Cell Line

Keywords

  • Air-liquid interface
  • Cryptosporidium parvum
  • Long-term growth
  • Primary cell
  • Transwell

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Wilke, G., Wang, Y., Ravindran, S., Stappenbeck, T., Witola, W. H., & Sibley, L. D. (2020). In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers. In Methods in Molecular Biology (pp. 351-372). (Methods in Molecular Biology; Vol. 2052). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9748-0_20

In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers. / Wilke, Georgia; Wang, Yi; Ravindran, Soumya; Stappenbeck, Thaddeus; Witola, William Harold; Sibley, L. David.

Methods in Molecular Biology. Humana Press Inc., 2020. p. 351-372 (Methods in Molecular Biology; Vol. 2052).

Research output: Chapter in Book/Report/Conference proceedingChapter

Wilke, G, Wang, Y, Ravindran, S, Stappenbeck, T, Witola, WH & Sibley, LD 2020, In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 2052, Humana Press Inc., pp. 351-372. https://doi.org/10.1007/978-1-4939-9748-0_20
Wilke G, Wang Y, Ravindran S, Stappenbeck T, Witola WH, Sibley LD. In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers. In Methods in Molecular Biology. Humana Press Inc. 2020. p. 351-372. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-9748-0_20
Wilke, Georgia ; Wang, Yi ; Ravindran, Soumya ; Stappenbeck, Thaddeus ; Witola, William Harold ; Sibley, L. David. / In Vitro Culture of Cryptosporidium parvum Using Stem Cell-Derived Intestinal Epithelial Monolayers. Methods in Molecular Biology. Humana Press Inc., 2020. pp. 351-372 (Methods in Molecular Biology).
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