Skip to main content
. 2018 May 22;7:e36491. doi: 10.7554/eLife.36491

Figure 4. H. hammondi can be successfully subcultured in vitro for a limited period of time.

(A) Monolayers containing HhCatAmer with oocyst debris were needle passaged, filtered, and used to infect a confluent monolayer of HFFs seven days post-excystation. Vacuoles were observed in subcultured monolayers after 6 DPI. (B–H) Confluent monolayers of HFFs were infected with HhCatAmer sporozoites at an MOI of ~2. (2,812,500 sporozoites). After a three-hour incubation at 37°C, the Day 0 transfer flask was scraped, needle passaged, filtered, and transferred to a new host cells. This process was repeated after 2, 5, 8, and 15 days of growth. Each flask was monitored daily for the number (C–H) for 13 DPI, or until visible vacuoles were no longer detected. The number of visible vacuoles increased in number for the first week post-excystation then began to decrease. Replicative capacity was greatest for sporozoites transferred to new host cells at Day 0 (C) and decreased during each subsequent passage. (D–H).

Figure 4.

Figure 4—figure supplement 1. H. hammondi vacuole sizes following subculture.

Figure 4—figure supplement 1.

(A–D) Visible vacuoles sizes were measured using ImageJ (NIH) for each day of attempted subculture. Vacuole size progressively increases over the course of in vitro cultivation, after which the size plateaus.