Why not use collagen typeI, fibronectin, and laminin5 in replacement for matrigel? (Ref; Metastasis Research Protocols Methods in Molecular Medicine™ Volume 58, 2001, pp 81-89) I also attach for you  the standard protocol for invasion assay.

Many researchers use the mixture of collagen-1 and matrigel for 3-D culture experiments. Further, tumor stiffness induces vessel collapse and makes hypoxic micro-environment especially in fibrosis-associated tumors such as pancreatic cancer. 

http://link.springer.com/protocol/10.1385%2F1-59259-137-X%3A081

Thanks Sir, 

Can i use any one like collagen type i or we have to use the mixture of collagen type I fibronectin and laminin5.

Can you send me the PDF of the protocol from springer .

Regards

For the invasion assays very nice alternative will be GeltrexTM (Life Technologies). For the references you can look for Rainero et al., J Cell Biol. 2012, 196(2):277-95. doi: 10.1083/jcb.201109112, and previous publications. They found that ovarian (A2780) and breast (MB-231) cancer cells invade best when the final concentration in the plug is between 6 and 8 mg/ml.

 I hope that will help.

I will try to prove my developed alternative within the next weeks/month and I hope it will work. I think it will! Please find www.TheHumanTouch.at for my ?crazy idea? to humanize cell culture - as good as I just can.

I am open for your opinion for my current research project. All the best for your research, Johannes