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Transwell Cell Migration/Invasion Assay


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4 replies to this topic

#1 cellcounter

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Posted 19 June 2009 - 11:16 AM

Hi,

Can anybody very well point out the differences in migration and invasion assays done using Transwell/Boyden chamber?

Specifically, what transwell chambers to use to study epithelial cell migration and invasion potential. It is important that you point out different protocols for measuring migration and that for measuring invasion separately. Ultimately, I need to present two graphs, one showing difference in migration rates for two cell lines and another showing difference in invasion rates between these two cell lines.

Any discussion on what matrix substance to use etc. would be greatly appreciated.

Thanks.

#2 pcrman

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Posted 19 June 2009 - 06:55 PM

Hi Cellcounter,

The following may answer your question of what is the difference between mirgation and invasion:

"Cell migration, the movement of cells from one area to another generally in response to a chemical signal, is central
to achieving functions such as wound repair, cell differentiation, embryonic development and the metastasis of tumors.
Cell invasion is similar to cell migration; however, it requires a cell to migrate through an extracellular matrix (ECM) or
basement membrane extract (BME) barrier by first enzymatically degrading the barrier in order to become established
in a new location. Cell invasion is exhibited by both normal cells in responses such as inflammation and by tumor cells
in the process of metastasis," from Corning protocol http://catalog2.corn...is_protocol.pdf

#3 cellcounter

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Posted 23 June 2009 - 07:54 AM

Hi Cellcounter,

The following may answer your question of what is the difference between mirgation and invasion:

"Cell migration, the movement of cells from one area to another generally in response to a chemical signal, is central
to achieving functions such as wound repair, cell differentiation, embryonic development and the metastasis of tumors.
Cell invasion is similar to cell migration; however, it requires a cell to migrate through an extracellular matrix (ECM) or
basement membrane extract (BME) barrier by first enzymatically degrading the barrier in order to become established
in a new location. Cell invasion is exhibited by both normal cells in responses such as inflammation and by tumor cells
in the process of metastasis," from Corning protocol http://catalog2.corn...is_protocol.pdf

Thanks pcrman!

#4 cancergeek

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Posted 27 June 2009 - 04:45 AM

There are a number of different platforms for measuring invasion and migration. For myself I use a transwell migration / invasion assay. By seeding cells in specially designed inserts (BD biosciences) with microscopic pores you can induce them to migrate by adding a chemoattractant such as high serum or high serum plus growth factors like IGF, PDGF or EGF to the well that the insert is placed into. The cells will then migrate through the pores in the insert membrane to the chemoattractant if they are capable of doing so. The addition of matrigel to the insert will allow you to also measure the cells ability to invade through the ECM. This system is very robust when dealing with small sample sizes or assessing drug treated versus non-treated controls. If you need something more high thoroughput there are HTS invasion or migration assays sold by companies such as Trevigen which allow you to assess invasion/migration through a BME on a 96-well plate using a fluorescence capable plate reader. The transwell assay is more well established currently and most of the literature you'll find will be based on that system. However the analysis can be somewhat tedious as membranes from each insert must be fixed, stained, cut out, mounted, photographed and counted.


Hope that helps.

Edited by cancergeek, 27 June 2009 - 05:59 AM.


#5 cellcounter

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Posted 29 June 2009 - 07:25 AM

There are a number of different platforms for measuring invasion and migration. For myself I use a transwell migration / invasion assay. By seeding cells in specially designed inserts (BD biosciences) with microscopic pores you can induce them to migrate by adding a chemoattractant such as high serum or high serum plus growth factors like IGF, PDGF or EGF to the well that the insert is placed into. The cells will then migrate through the pores in the insert membrane to the chemoattractant if they are capable of doing so. The addition of matrigel to the insert will allow you to also measure the cells ability to invade through the ECM. This system is very robust when dealing with small sample sizes or assessing drug treated versus non-treated controls. If you need something more high thoroughput there are HTS invasion or migration assays sold by companies such as Trevigen which allow you to assess invasion/migration through a BME on a 96-well plate using a fluorescence capable plate reader. The transwell assay is more well established currently and most of the literature you'll find will be based on that system. However the analysis can be somewhat tedious as membranes from each insert must be fixed, stained, cut out, mounted, photographed and counted.


Hope that helps.

Hi cancergeek,

Thanks for your detailed reply. In fact, after studying the various options, I had purchased Trevigen (from R&D) kit with 96-well format. As you pointed out, fluorescence should reduce the many variables associated with staining, counting etc in the transwell assay.

It turns out a bit more expensive, but I hope less time consuming and reproducible! It is also a more compact kit--without necessity to pool reagents from various companies.




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