Larvae, Pupae, and Vero Cells

Earlier this week, we hatched our eggs, separated them into groups of 250 larvae per tray, fed them, and let them develop. After sorting all the larvae, we ended up with 24 trays of each mosquito population resulting in a grand total of 12,000 mosquitoes! These pictures are all of the larval rearing trays that we currently have in the rearing room.

Under ideal temperature conditions and with adequate food, this is about the time we should start seeing pupae. Sure enough, when I came to the lab this morning, I was greeted by hundreds and hundreds of pupae! A good portion of the morning was spent transferring the pupae from the larval rearing trays into small cups. These cups were then placed in a cage with sugar water. Within the next 1-2 days, these pupae will be adult mosquitoes.

This weekend, in preparation for the experiment, we also had to seed our flasks with Vero cells. Vero cells are a lineage of cells that are used in cell culture and these are cells that we will eventually infect with our virus. However, before we can infect the cells, we must first know how many cells are in each flask. This is where the process of seeding comes in. Seeding is spreading a defined amount of cells into the flask. The cells that we distribute in the new flask are pulled from an existing cell culture. After a few steps, the cells are dislodged and present in a cell suspension (video below – ignore the mosquito stuck somewhere in the microscope). From here, we calculate how much cell suspension is needed to get 21.6×10^6 cells per flask, add some media, and allow the cells to incubate for a few more days.

The major task this week will be picking pupae, but on Thursday, Friday, and Saturday, we will also be inoculating our cells with Zika virus. One step closer to our infectious bloodfeeding!

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