20 February 2019 5 9K Report

Hello,

Recently I performed lentiviral transduction on several cell lines. On day one with the lentivirus, I observed black aggregates moving (dancing, vibrating) around my cells. These are only in cells to which I added virus.

When I harvested my lentivirus in the supernatant of my packaging cell line, I spun the supernatant (3000RPM for 15 minutes) to remove cellular debris, filtered it with a 450nm sterile filter, and then froze it at -80C. These steps should have removed bacteria, correct? Moreover, the aggregates are resistant to the 1% Pen/Strep in my culture media. They do not turn the media turbid, and they never seem to reach log phase.

Could I be seeing the virus itself?

Can lentiviruses form visible aggregates?

Could these be abiotic aggregates exerting Brownian motion?

I have seen this phenomenon before and as of now, no one has given me a satisfactory answer regarding the identity of the dots.

Thank you.

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