4 replies to this topic

[Vera9]

Hi everybody,

I'd be very pleased if someone could tell me (or give some links where I can find the info) the difference between Transwell or Boyden Chamber assay and the Scratch Wound / Wound healing assay.

The thing is that I've done some experiments with an inhibitor and I get different results in Transwell assay and in Scratch Wound. In Transwell I see an increase in cell migration whereas in SWA I don't see any effect. Transwell assay measures the ability of cells to migrate towards a chemoatractant, is that right? What kind of migration does SWA analize?

Thanks in advance for any help.

Vera9

[Inmost sun]

Vera9, on Jul 15 2010, 03:26 PM, said:

Hi everybody,

I'd be very pleased if someone could tell me (or give some links where I can find the info) the difference between Transwell or Boyden Chamber assay and the Scratch Wound / Wound healing assay.

The thing is that I've done some experiments with an inhibitor and I get different results in Transwell assay and in Scratch Wound. In Transwell I see an increase in cell migration whereas in SWA I don't see any effect. Transwell assay measures the ability of cells to migrate towards a chemoatractant, is that right? What kind of migration does SWA analize?

Thanks in advance for any help.

Vera9

In both assays you can measure dividing and/or migrating cells; with scratch wound assay you measure the ability of a tissue-like situation to close a pertubation of integrity; you detect cell migration in a transwell experiment but don´t you really detect any cell migration or cell division in in a scratch wound assay?

[Vera9]

Hi Inmost sun and thanks for your answer.

I detect migration in transwell assay, and it is increased when I treat the cells with the inhibitor. But when doing SWA, I can't see differences between the control and the inhibitor treated cells, they close the wound aproximately at the same time.

I've also seen that this inhibitor stops cell cycle at G1/S. Maybe what I'm seeing in SWA is a mixture of the effects of the increase in migration and cell cycle arrest. I mean, maybe when I only look at migration (like in transwell) I see the increase, but when looking both at migration and proliferation (like in SWA), one thing compensates the other so I see no effect... could that be possible?

Thanks in advance!

Vera9

[Inmost sun]

Vera9, on Jul 19 2010, 12:32 PM, said:

Hi Inmost sun and thanks for your answer.

I detect migration in transwell assay, and it is increased when I treat the cells with the inhibitor. But when doing SWA, I can't see differences between the control and the inhibitor treated cells, they close the wound aproximately at the same time.

I've also seen that this inhibitor stops cell cycle at G1/S. Maybe what I'm seeing in SWA is a mixture of the effects of the increase in migration and cell cycle arrest. I mean, maybe when I only look at migration (like in transwell) I see the increase, but when looking both at migration and proliferation (like in SWA), one thing compensates the other so I see no effect... could that be possible?

Thanks in advance!

Vera9

so, you may perform additionally proliferation assays (MTT, SRB, etc); I would try to analyze the SWA dynamically f.i. in video microscopy; you will better discriminate between diving of cells and cell migration

[Vera9]

Hi Again,

I've already done MTT and SRB.

I'll try what you say, monitorize the SWA. Hope i'll be able to see something.

Thanks.