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Long-term storage of 'Cryptosporidium parvum' for in vitro culture

Paziewska-Harris, A., Schoone, G. and Schallig, H. D. F. H. 2018. Long-term storage of 'Cryptosporidium parvum' for in vitro culture. Journal of Parasitology 104 (1) , p. 96. 10.1645/16-22

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Abstract

The long-term storage of Cryptosporidium life-cycle stages is a prerequisite for in vitro culture of the parasite. Cryptosporidium parvum oocysts, sporozoites, and intracellular forms inside infected host cells were stored for 6–12 mo in liquid nitrogen utilizing different cryoprotectants (dimethyl sulfoxide [DMSO], glycerol and fetal calf serum [FCS]), then cultured in vitro. Performance in vitro was quantified by estimating the total Cryptosporidium copy number with quantitative polymerase chain reaction (qPCR) in 3- and 7-day-old cultures. Although few parasites were recovered either from stored oocysts or from infected host cells, sporozoites stored in liquid nitrogen recovered from freezing successfully. More copies of parasite DNA were obtained from culturing those sporozoites than sporozoites excysted from oocysts kept at 4 C for the same period. The best performance was observed for sporozoites stored in Roswell Park Memorial Institute (RPMI) medium with 10% FCS and 5% DMSO, which generated 240% and 330% greater number of parasite DNA copies (on days 3 and 7 post-infection, respectively) compared to controls. Storage of sporozoites in liquid nitrogen is more effective than oocyst storage at 4 C and represents a more consistent approach for storage of viable infective Cryptosporidium aliquots for in vitro culture.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: American Society of Parasitologists
ISSN: 0022-3395
Date of First Compliant Deposit: 12 April 2019
Date of Acceptance: 2 November 2017
Last Modified: 01 Jul 2019 11:55
URI: http://orca-mwe.cf.ac.uk/id/eprint/121739

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