Infectious Rates of HCT-8 Cells Infected with Cryptosporidium Parvum Sporozoites Obtained in In Vitro Different Excystation Conditions

ABSTRACT

In this study, HCT-8 cells infected with C. parvum sporozoites which were released in 3 different excystation conditions (TA/cytomixs:RPMI-1640 excystation media (1:1); TA/cytomix:RPMI-1640 media and TA/cytomix media) were counted under fluorescent microscopy by IFA (Immunofluorescent assay method).

Different pulses (pulse 1, pulse 2, pulse 3 and heat inactivated) were applied for C. parvum sporozoities under electroporation conditions of 1000 voltage (kV/cm), 50 capacitance (μF) and 50 recistance (ohm Ω) and cell monolayers were counted under fluorescent microscopy to determine infected cells. As a consequence, the maximum infected cell monolayers were enumerated at pulse 1 under electroporation conditions of 1000V, 50μ 50Ω

Our results showed that the best condition of excystation was TA/cytomixs:RPMI-1640 excystation media (1:1) with excystation rate of 80% and the other excystation rates were 70% and 60% for TA/cytomix:RPMI-1640 media and TA/cytomix media, respectively.

Keyword:

• HCT-8 cells culture
• electroporation
• excystation
• C. parvum
• immunofluorescence assay (IFA)