3D culture of colorectal cells- suggestion please!

6 posts / 0 new
Last post
vdas
vdas's picture
3D culture of colorectal cells- suggestion please!

Hello people,

Seeking help. 
So I am starting this new project on 3D culture, and practically have no knowledge about 3D culture system. To begin with, I'm (we are) planning to culture colorectal cell lines (HTC116) in 3D, and then move to primary culture of colorectal cells in 3D. I need a simple and cost-effective system to present some positive data to our director and get a nod for the direction  I am thinking to take for the project. I wanted to know if it is possible to grow these cells into spheroids in agar coated plates or matrigel? If not, please suggest some alternatives. Are there any other method that could be easily established in the lab without having to spend heaps just in the beginning? 
I read about breast cancer cell lines that can be easily grown in 3D and they start expressing tumor-like charateristics. Do only particular cells have this ability to form spheriod or any adherent cell line will form spheroids under suitable conditions? A
Lastly, is it possible to do MTT on spheriods grown in a 96-well plate to determine IC50 values?
Thanks a lot

Jason King
Jason King's picture
 A cheap method is to put the

 A cheap method is to put the cells into 96 well plates then invert the lates to generate hanging drops. If you're lucky they will form spheroid a on their own. If not you might stimulate this by including some collagen with the cells at the beginning. I would do a Google search for HTC116 AND "hanging drop" as it's almost certainly been done before. No point re-inventing the wheel.

Cfroberg
Cfroberg's picture
 Check out the n3D Bio cell

 Check out the n3D Bio cell culture system.  It is as easy as 2D, inexpensive and very fast.  They will set up a webinar for you to go over the system and applications.  Check out the wiki on 3D cell culture via magnetic levitation or www.n3dbio.com.

vdas
vdas's picture
 Thank you. Will do the

 Thank you. Will do the Google search and also look into n3D culture system.

vdas
vdas's picture
 Just posting an update-

 Just posting an update-
Hanging drop works perfectly fine. It generates perfect spheroids with consistent volume. However, the process is quite laborious to incorporate into a 384-well plate for HTS. I have found agarose-coating method quite useful for a 384-well plate. 

Synthecon 3D
Synthecon 3D's picture
You should check out this 3D

You should check out this 3D cell culture technology: http://www.synthecon.com/company/technology.html

Here are a few sample papers: http://www.synthecon.com/cancer.html

and one directly relevant to you: http://cancerres.aacrjournals.org/content/64/8/2643.long

This system generates more viable and biomimetic spheroids than any other technology. They have FAQs and training to help you get started.