Hello All,
I am trying to do a 3D cell culture for Smooth Muscle cells and Vascular Endothelial Cell lines.
I found a lot of reviews and publications but I was wondering if anyone can refer me to a step-wise protocol for culturing the cells either using natural or synthetic hydrogels. (not the suspension systems)
Thank you
Hi, Maedeh.
You seems to want the procedure to be followed without thinking and challenge all the aspects of 3D culture system, including cells, what tissues, what gels, and pitfalls and limitations. The more you read, the smarter you become as you think about lots of things. Don't give up such precious challenges.
However, if you want such a protocol to be followed simply without thinking, there is one (link). http://bio-protocol.org/e2247
Good luck!
Hello Maedeh
Given what facilities available in your lab and what you are planing to do It is not easy to find a exact match protocol, but you always can customize your method/protocol. for example we while had a 3D system consisted of a custom designed bioreactor, media reservoir, peristaltic pump and tubing. the bioreactor was a chamber to house the scaffold and designed to allow laminar flow. The growth chamber that housed the scaffold was few mm in diameter to house scaffolds. The growth chamber was surrounded on both sides by a metal mesh with notches to allow air bubbles to accumulate and avoid the scaffold. generally nothing complicated.
The point is see what you have and consult good books and set uo your system. papers normally are precise and hard to get details out of them
Technology Platforms for 3D Cell Culture: A User's Guide
3D Cell Culture: Methods and Protocols
I hope it will help
Amin
Hi, Maedeh.
You seems to want the procedure to be followed without thinking and challenge all the aspects of 3D culture system, including cells, what tissues, what gels, and pitfalls and limitations. The more you read, the smarter you become as you think about lots of things. Don't give up such precious challenges.
However, if you want such a protocol to be followed simply without thinking, there is one (link). http://bio-protocol.org/e2247
Good luck!
Thank you Dr. Lee for your insights.
I 100% agree with you, though 3D cell culture is a very small portion of the project that I am doing and that's why I'm not going to be focusing on them too much. We are to design a miniaturized optical biosensor to be input in a microfluidic organ on a chip device and 3D cell culture is just a basic process and that's why I didn't want to spend too much time on specializing in it. I have read the generals, of course, but going deep in that area will steal my focus from the original design.
Thank you so very much for the heads up, and yes, I completely agree that we always have to keep our brains at 20% workload :)
Thank you Amin. Your set up sounds very creative and interesting.
Guess I'll end up designing one as well. Thank you.
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