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?

Post  Anirban on Tue Jan 27, 2009 4:18 pm

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

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

Post  ThomasMCS on Wed Jan 28, 2009 8:36 pm

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

Post  Richard2014 on Fri Jan 10, 2014 5:19 am

If MEAs are treated with (for example) fetal bovine serum (for 3 days, 4C), 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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Re: Is it possible to make a new MEA hydrophilic without plasma-cleaning/protein coating?

Post  Thomas on Sun Jan 12, 2014 9:15 pm

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

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

Post  Richard2014 on Thu Jan 23, 2014 11:25 pm

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

Have a nice weekend,
Richard

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

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