Preparing 3D Liver Spheroids

Preparation of in vitro 3D liver spheroid models in hanging-drop formation. Spheroids are then applied to micronucleus analysis through the Cytokinesis Blocked Micronucleus Assay.
In Vitro Toxicology Group, Swansea University, Wales, U.K.
Lab work performed by Samantha Llewellyn
Filmed by Demi Pritchard
Edited and narrated by Dr Kate Chapman

[@rupeshthorat3058]

Thanks for the detailed information. This video is awesome

[@NikhilMandrekar]

Thanks, it would be nice to explain the "why" behind each step (for the average person)

[@Indresh2468]

Thanks for this video! I will try to implement this technique to generate mammospheres.

[@vahidfallah8730]

Thank you for the clear explanation, I also want to use this method in my experiment, do you know how can I transfect the 3D culture cells with Lenti Virus?

[@jaydebpal1818]

Hello ma'am,
Can you please upload the procedure for spheroid generation in 6 well plates in ultra low attachment plates for Huh7 cell lines

[@srijanidey6240]

Hello ma'am,
I have tried to replicate the process as described in the video and encountered some problems :
1. loss of drops in PBS solution while taking out of incubator
2. Despite incubating the drops for 4 days, I failed to get a pellet of organoid. (I have centrifuged at 100rpm for 5 mins)
3. Was unable to observe the progression of organoids in a microscope.
I will be highly obliged if you kindly share your opinion regarding these.

[@swethakrishnamani9632]

Can this be done when we use hydrogel

[@neelhinsu7971]

Hello mam, can you give me detailed protocol after spheroids are formed by hanging drop method!?

[@user-ro8ou8fm8o]

What is the rpm and time duration at which the spheroids were centrifuged during the harvesting procedure?

[@kosheeka]

In most cases, centrifugation at 200 x g for 5 minutes is sufficient to pellet cells following trypsinization. However, if the culture medium is highly viscous, you might need to extend the centrifugation time. Five minutes remains a good starting point, but a slight increase may be necessary for more viscous culture mediums.

[@barbaradiasbiomed]

Is this a specific plate for hanging drops?

[@Indresh2468]

You could use a normal 60 or 100mm petridish. It works just as well.