Hello.

What kind of technique do you use to create your spheroids? And what is their average size?

If you use a technique where each spheroid grows within a unique well, eg hanging drop, then when in pproper size (depending on the type of your cells), they can be isolated simply by using a pipette. If your spheroid remains compact after transfer, then it could be either parafin-embedded or cut using a microtome?

-Marilena- 

I think Elplasia will help your experiments. Please refer to Elplasia website.

http://www.elplasia.com/products/

Hi, Thank you all for your useful answers.

I am using agarose gel in 96-well plate technique for HepG2 (liver cancer) and MDA-MB-231 (breast cancer) cell lines. But I need to transfer spheroids to 24-well plate after 4-6 days; then can start the treatment.

Regards;

Pooya

I also use MDA. They grow up perfect in Matrigel instead of agarose. No attachment in well is observed.

-Marilena-

Hi,

I am planning to do some experiments with tumor spheroids. I have the same question like Pooya (question 2). How can I wash the spheroids with PBS after transferring them into another plate? 

(As the spheroids are not attached with the plates, they can be washed away with PBS) 

Carefully use a 10ul tip (white) to wash them with PBS. You may rather do this procedural step in a cverslide and afterwards use a larger diameter tip to transfer the spheroid to the plate.

In my experience, MB - 231s and MCF - 7 s do not consistently form compact spheroids by hanging drop method until some matrigel, agar based gel or Poly(N-isopropylacrylamide) is added to the media while seeding the spheroids. More adherent cells like 4 T1 form good spheroids even without adding any gel by hanging drop method.