Cardiomyocytes culture in MEA

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Cardiomyocytes culture in MEA

Post  mcs on Thu Jun 12, 2008 9:44 am

We made cultures of cardiomyocytes of neonatal rat in the MEA. The cells do not beat. We used collagen for coating substrat. Does somebody have ideas?

mcs

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Re: Cardiomyocytes culture in MEA

Post  mcs on Thu Jun 12, 2008 9:44 am

Dear Sabir,

the collagene coating should be fine. However we use Nitrocellulose or polyethylenimimne.

Can you give us some more details regarding your preparation ?
- How old are the animals ?
- what culture media do you use ?
- how dense do you plate the cells ?
- Do they beat when you plate them on a culture dish ?

Even so you use rat and not chicken, I would recomend to have a look on our application note regarding the chicken embryo preparation where we give some general hints for cardiomyocyte recording on the MEA

http://www.multichannelsystems.com/support/applicationnotes.htm

Greetings

Thomas

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Re: Cardiomyocytes culture in MEA

Post  mcs on Thu Jun 12, 2008 9:44 am

Dear Thomas,

The animals are neonatal rats (2 to 4 days old).
We use a ventricular media.
We plate between 0.5*10^6 and 0.7*10^6 cells.
We don't say if cells beat when we plate them on a culture dish because they are in a cell solution.

Thanks,

sabir

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Re: Cardiomyocytes culture in MEA

Post  mcs on Thu Jun 12, 2008 9:45 am

Dear Sabir,

the concentration and the animal age sound good.

Some little hints which helped us to improve beating efficiency :
- we plate one little 10 ul drop directly on the recording area of the
MEA, let the cells attach for about one minute before we fill up with
medium. Thus we receive a very high local density
- We reduce the FCS concentration in the medium to 3% which yielded
higher percentages of beating cells

Greetings

Thomas Meyer

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Re: Cardiomyocytes culture in MEA

Post  mcs on Thu Jun 12, 2008 9:45 am

Dear Thomas,
A few more Qs: (a)how do you use a nitrocellulose coating? Do you dissolve NC in MeOH and then coat coverslips? (b) Ventricular media?
This is not for the MEA assay but for the tissue culture experiment.
Thanx,
Vicki

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Re: Cardiomyocytes culture in MEA

Post  mcs on Thu Jun 12, 2008 9:46 am

Dear Vicki,

please have a look in our dowmloadsection as well.
There you will find application notes for cardiomyocyte culture as well as general MEA handling tips covering coating ...

http://www.multichannelsystems.com/support/applicationnotes.htm

I dissolve the NC membrane in MetOH and coat as describe above.
For detailed Media composition please see our application note

Greetings

Thomas

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