Convenient Protocol for Production and Purification of Clostridioides difficile Spores for Germination Studies

Melissa Weldy; Clayton Evert; Peter I Dosa; Alexander Khoruts; Michael J Sadowsky

doi:10.1016/j.xpro.2020.100071

Convenient Protocol for Production and Purification of Clostridioides difficile Spores for Germination Studies

STAR Protoc. 2020 Jul 24;1(2):100071. doi: 10.1016/j.xpro.2020.100071. eCollection 2020 Sep 18.

Authors

Melissa Weldy^{1

2}, Clayton Evert^{1

2}, Peter I Dosa³, Alexander Khoruts^{1

2}, Michael J Sadowsky^{2

4}

Affiliations

¹ Department of Medicine, Division of Gastroenterology, University of Minnesota, Minneapolis, MN 55414, USA.
² BioTechnology Institute, University of Minnesota, St. Paul, MN 55108, USA.
³ Institute for Therapeutics Discovery and Development, Department of Medicinal Chemistry, University of Minnesota, Minneapolis, MN 55414, USA.
⁴ Departments of Plant & Microbial Biology, and Soil, Water & Climate, University of Minnesota, St. Paul, MN 55108, USA.

Abstract

Clostridioides difficile, an obligate anaerobic bacterium, causes infections leading to prolonged diarrhea. The bacterium produces dormant spores that can withstand an aerobic environment, resulting in easy environmental transfer. Here, we present a convenient sporulation and purification protocol that can be practiced in any lab setting using a portable anaerobic glove bag. This protocol also optimizes existing cell growth methods and presents a detailed trouble shooting guide. This protocol is a modification of those previously reported by Edwards and McBride (2016) and Shen et al. (2016).

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Bacteriological Techniques / methods*
Cell Culture Techniques / methods*
Clostridioides difficile* / cytology
Clostridioides difficile* / metabolism
Spores, Bacterial* / cytology
Spores, Bacterial* / isolation & purification
Spores, Bacterial* / metabolism

Grants and funding

U01 HL127479/HL/NHLBI NIH HHS/United States