Migration -or rather non-migration- of smooth muscle cells - (Sep/06/2005 )

Hello everybody!

We´d like to try out an migration assay with human smooth muscle cells (HCASMC), but the cells don´t even think of attaching to the membrane!

We use Nunc cell culture inserts with a polycarbonate membrane with a pore size of 8 µm. In my first attempt i used the inserts "untreated", only unpacked. In the second i coated the membranes with collagen. In both tries the cells didn´t attach to the membrane.

So, my question: has anyone some more effective experience with such inserts? Why do the cells not attach to the membrane?

I´m curious about (hopefully) numerous answers!

Anja

Hi, how is the situation right now?
I have the same problem . Using Corning transwell, the SMCs never attach to the insert, even after treated with collagenI or poly-L-glycin.
Don't know what to do next.

Hi!

I would like to ask you whether in the meanwhile you had success in establishing a migration assay for VMSCs, or whether anybody else has any ideas about troubleshooting of a VSMC-cell migration assay?

I'm working with rat VSMC and I don't get the cells to migrate towards PDFG, with and without collagenI, they simply stay on top. so it's not possible to go any further with my experiments (testing the anti-migration-potential of my substance) :-(

The inserts (I'm working with BD, Falcon inserts, 8uM) are very expensive, and I already wasted a lot of them, I changed all parameters (time, amount of collagen, amount of PDGF, amount of cells, passage of cells, starving the cells 24h before seeding, not starving the cells, ...).

I would be very grateful for any hints/ protocols!!!


Irene

Hello everybody!

We´d like to try out an migration assay with human smooth muscle cells (HCASMC), but the cells don´t even think of attaching to the membrane!

We use Nunc cell culture inserts with a polycarbonate membrane with a pore size of 8 µm. In my first attempt i used the inserts "untreated", only unpacked. In the second i coated the membranes with collagen. In both tries the cells didn´t attach to the membrane.

So, my question: has anyone some more effective experience with such inserts? Why do the cells not attach to the membrane?

I´m curious about (hopefully) numerous answers!

Anja

Can you seed SMC on the plate, create a scratch, wait and see if SMCs migrate and fix the gap?