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Is it possible to make a new MEA hydrophilic without plasma-cleaning/protein coating?

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Is it possible to make a new MEA hydrophilic without plasma-cleaning/protein coating? Empty Is it possible to make a new MEA hydrophilic without plasma-cleaning/protein coating?

Post  Anirban Wed Jan 28, 2009 4:18 am

Hi
Is there any way to make a new MEA (200/30 int. ref.) hydrophilic other than by plasma-cleaning or coating with a protein like albumin? I do plan to use PEI/PDL and Laminin coating prior to culturing the cells, but am guessing these will not coat uniformly without some prior treatment of the MEA. Thanks for your time and help.
Ani

Anirban

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Post  ThomasMCS Thu Jan 29, 2009 8:36 am

We use the plasma cleaner in our laboratory to hydrophilize the MEAs. This works nicely and quite reproducible.
As you mentioned an alternative is working with high proteins solutions, such as FCS. This works, however it is less standardized and not so reproducable.

However I am not aware of other methods than these two. Is there a reason why you are looking for alternatives? Are the standard methods not working for you?

Greetings

Thomas

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Post  Richard2014 Fri Jan 10, 2014 5:19 pm

If MEAs are treated with (for example) fetal bovine serum (for 3 days, 4°C), how should be proceeded before PDL coating take place?

Should we flush them with PBS?
Or just aspirate the (1ml) of FBS on a standard-MEA and coat them with PDL directly after removing FBS?

Thanks!

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Post  Thomas Mon Jan 13, 2014 9:15 am

Rinse it briefly with PBS, however, our preferred prodeure is plasma treatment: faster, more efficeint and more reproducible.
Thomas

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Post  Richard2014 Fri Jan 24, 2014 11:25 am

Thanks for your advise!

We will check if we can get a plasma cleaning device for our institution.


So far, the FBS-treatment was sufficient for our sowing two weeks ago. Cells on MEAs, treated with FBS before the application of PDL, followed by Laminin, attached pretty well to the surface of the MEAs; while FBS untreated MEAs had a higher loss rate.

Our protocol:

1. Autoclave MEAs.
2. Pipett 50µl of FBS on the electrode field only and store standard-MEAs for 4 days on 4°C.
3. Aspirate FBS.
4. Wash MEA with 1ml of PBS two times.
5. Let MEAs dry under the bench.
5. 50µl PDL directly on the electrode field and store them over night (4°C).
6. Aspirate PDL, let them dry.
7. 50µl Laminin directly on the electrode field and store them for 3-4 days (4°C) before the attachment of cells.

Have a nice weekend,
Richard

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