Scalable cryopreservation of infectious Cryptosporidium hominis oocysts by vitrification

Justyna J. Jaskiewicz; Denise Ann E. Dayao; Donald Girouard; Derin Sevenler; Giovanni Widmer; Mehmet Toner; Saul Tzipori; Rebecca D. Sandlin

doi:10.1371/journal.ppat.1011425

Scalable cryopreservation of infectious Cryptosporidium hominis oocysts by vitrification

Justyna J. Jaskiewicz, Denise Ann E. Dayao, Donald Girouard, Derin Sevenler, Giovanni Widmer, Mehmet Toner, Saul Tzipori, Rebecca D. Sandlin

Advanced Technologies for Preservation of Biological Systems

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3 Scopus citations

Abstract

Cryptosporidium hominis is a serious cause of childhood diarrhea in developing countries. The development of therapeutics is impeded by major technical roadblocks including lack of cryopreservation and simple culturing methods. This impacts the availability of optimized/ standardized singular sources of infectious parasite oocysts for research and human challenge studies. The human C. hominis TU502 isolate is currently propagated in gnotobiotic piglets in only one laboratory, which limits access to oocysts. Streamlined cryopreservation could enable creation of a biobank to serve as an oocyst source for research and distribution to other investigators requiring C. hominis. Here, we report cryopreservation of C. hominis TU502 oocysts by vitrification using specially designed specimen containers scaled to 100 μL volume. Thawed oocysts exhibit ~70% viability with robust excystation and 100% infection rate in gnotobiotic piglets. The availability of optimized/standardized sources of oocysts may streamline drug and vaccine evaluation by enabling wider access to biological specimens.

Original language	English (US)
Article number	e1011425
Journal	PLoS pathogens
Volume	19
Issue number	6
DOIs	https://doi.org/10.1371/journal.ppat.1011425
State	Published - Jun 2023

Bibliographical note

Funding Information:
The research was funded by the National Institutes of Health award R21AI154026 granted to R.D.S. and S.T., the National Science Foundation award EEC 1941543 granted to M.T. and the Good Ventures Foundation award granted to S.T. Support from R21AI154026 was received by J.J., R.S, M.T., G.W., and S.T., from EEC 1941543 by J. J., D.S., M.T. and R.S., and from Good Ventures Foundation award by S.T. and D.D. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors acknowledge Jennipher Grudzien from Grace Bio-Labs for help with fabrication of the high-aspect ratio cassette.

Publisher Copyright:
© 2023 Jaskiewicz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PubMed: MeSH publication types

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

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title = "Scalable cryopreservation of infectious Cryptosporidium hominis oocysts by vitrification",

abstract = "Cryptosporidium hominis is a serious cause of childhood diarrhea in developing countries. The development of therapeutics is impeded by major technical roadblocks including lack of cryopreservation and simple culturing methods. This impacts the availability of optimized/ standardized singular sources of infectious parasite oocysts for research and human challenge studies. The human C. hominis TU502 isolate is currently propagated in gnotobiotic piglets in only one laboratory, which limits access to oocysts. Streamlined cryopreservation could enable creation of a biobank to serve as an oocyst source for research and distribution to other investigators requiring C. hominis. Here, we report cryopreservation of C. hominis TU502 oocysts by vitrification using specially designed specimen containers scaled to 100 μL volume. Thawed oocysts exhibit ~70% viability with robust excystation and 100% infection rate in gnotobiotic piglets. The availability of optimized/standardized sources of oocysts may streamline drug and vaccine evaluation by enabling wider access to biological specimens.",

author = "Jaskiewicz, {Justyna J.} and Dayao, {Denise Ann E.} and Donald Girouard and Derin Sevenler and Giovanni Widmer and Mehmet Toner and Saul Tzipori and Sandlin, {Rebecca D.}",

note = "Funding Information: The research was funded by the National Institutes of Health award R21AI154026 granted to R.D.S. and S.T., the National Science Foundation award EEC 1941543 granted to M.T. and the Good Ventures Foundation award granted to S.T. Support from R21AI154026 was received by J.J., R.S, M.T., G.W., and S.T., from EEC 1941543 by J. J., D.S., M.T. and R.S., and from Good Ventures Foundation award by S.T. and D.D. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors acknowledge Jennipher Grudzien from Grace Bio-Labs for help with fabrication of the high-aspect ratio cassette. Publisher Copyright: {\textcopyright} 2023 Jaskiewicz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

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AU - Sevenler, Derin

AU - Widmer, Giovanni

AU - Toner, Mehmet

AU - Tzipori, Saul

AU - Sandlin, Rebecca D.

N1 - Funding Information: The research was funded by the National Institutes of Health award R21AI154026 granted to R.D.S. and S.T., the National Science Foundation award EEC 1941543 granted to M.T. and the Good Ventures Foundation award granted to S.T. Support from R21AI154026 was received by J.J., R.S, M.T., G.W., and S.T., from EEC 1941543 by J. J., D.S., M.T. and R.S., and from Good Ventures Foundation award by S.T. and D.D. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors acknowledge Jennipher Grudzien from Grace Bio-Labs for help with fabrication of the high-aspect ratio cassette. Publisher Copyright: © 2023 Jaskiewicz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Scalable cryopreservation of infectious Cryptosporidium hominis oocysts by vitrification

Abstract

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