Matrigel

I heard that matrigel is used to study matrix degradation in cancer cell invasion assays. What is matrigel?

Does anyone have a protocol for the Invasion assay?

Becton Dickson sells Matrigel. To quote from their website...
"BD Matrigel Matrix is a solubulized basement membrane preparation extracted from EHS mouse sarcoma, a tumor rich in ECM proteins. Its major component is laminin, followed by collagen IV, heparan sulfate proteoglycans, and entactin 1. At room temperature, BD Matrigel Matrix polymerizes to produce biologically active matrix material resembling the mammalian cellular basement membrane. Cells behave as they do in vivo when they are cultured on BD Matrigel Matrix. It provides a physiologically relevant environment for studies of cell morphology, biochemical function, migration or invasion, and gene expression."

You may want to consult their website for further information
BD Matrigel Information

Edited to fix url.

I hope this helps.

You can also check this protocol on the is site for plating in matrigel. Edited to add protocol.

Matrigel plating protocol

Have you managed to get hold of a protocol yet? I worked in an institute where one of the groups did invasion assays quite frequently. I can put you in touch if you'd like.Just drop me a private message.

I found this on www.protocol-online.org:

Matrigel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens, laminin, and proteoglycans)but also matrix degrading enzymes/their inhibitors and growth factors. Invasion of tumor cells into Matrigel has been used to characterize involvement of ECM receptors and matrix degrading enzymes which play roles in tumor progression.


Procedure
1. Thaw Matrigel at 4C overnight.
2. Dilute Matrigel (5mg/ml to 1 mg/ml) in serum free-cold cell culture media (RPMI1640, EMEM, DMEM, etc).
3. Put 100 ul of the diluted matrigel into upper chamber of 24-well transwell
4. Incubate the transwell at 37C at least 4 to 5 h for gelling.
5. Harvest cells from tissue culture flasks by Trypsin/EDTA.
6. Wash the cells 3 times with culture media (RPMI1640, EMEM, DMEM etc)containing 1 % FBS.
7. Resuspend the cells in media containing 1% FBS at a density of 10^6 cells/ml.
8. Gently wash gelled matrigel with warmed serum free-culture media.
9. Put 100 ul of the cell suspension onto the matrigel.
10. lower chamber of the transwell is filled with 600 ul of culture media containing 5 ug/ml fibronectin, as an adhesive subtrate.
11. Incubate at 37C for 20 to 24 h.
12. Remove transwells from 24-well plates and stained with Diff-Quick solution.
13. Scrape off noninvaded cells on the top of the transwell with a cotton swab.
14. Count invaded cells under a light microscope.

Recipes
- Matrigel (Becton-dickinson)
- 24-transwell (Coster)
- Fibronectin(Sigma)
- Diff-Quick staining solution (Fischer Scientific)

BD Biosciences does:
Link

Sigma has one:
http://www.sigmaaldrich.com/catalog/search/ProductDetail/SIGMA/E1270

But how do you use the Matrigel for 3D cell cultures. How do you handle it. Like, how do you make it liquid to make transfer into wells possible??

Hi,

I bought the BD Matrigel # 356231. I want to grow my cells in 3D resuspension. However, I cannot find any information about the concentration of this product and how much I can dilute it with my culture medium and it will still form 3D-gel. Do you know? Have you tried this?
Thank you very much!
x

The concentration differs from batch to batch. You have to call BD and give them the lot # to get the concentration of a particular batch.

Here's a paper with thorough description of the method for cell culture in 3D matrigel.

Methods
Volume 30, Issue 3 , July 2003, Pages 256-268

This is the pubmed link http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12798140&query_hl=8

Good luck!

[quote=xzhao]Hi,

I bought the BD Matrigel # 356231. I want to grow my cells in 3D resuspension. However, I cannot find any information about the concentration of this product and how much I can dilute it with my culture medium and it will still form 3D-gel. Do you know? Have you tried this?
Thank you very much!
x

Does anyone know the approximate concentration of matrigel as sold by BD. Neither Sigma nor BD will tell me, and it's pretty hard to design an experiment without a concentration.


[quote=shard602]The concentration differs from batch to batch. You have to call BD and give them the lot # to get the concentration of a particular batch.

Here's a paper with thorough description of the method for cell culture in 3D matrigel.

Methods
Volume 30, Issue 3 , July 2003, Pages 256-268

This is the pubmed link http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=12798140&query_hl=8

Good luck!

[quote=xzhao]Hi,

I bought the BD Matrigel # 356231. I want to grow my cells in 3D resuspension. However, I cannot find any information about the concentration of this product and how much I can dilute it with my culture medium and it will still form 3D-gel. Do you know? Have you tried this?
Thank you very much!
x

[/quote]

does anyone know the complete list of components in Matrigel? does it have glucose in it or is that only introduced if you mix it with growth medium? Thanks!