I have done this with mouse ear punches and it is pretty straightforward. I tried tails but it wasn't as easy. The fibros will actually migrate out of the ear/tail pieces and cover the dish. You basically just chop the ear with a razor blade, put it in trypsin, then plate it in media.

In a few days, you just pick out the ear pieces and all you are left with is fibros.

I will say it can take a 3-7 days so don't give up on the culture too early!

This website has a protocol from Current Protocols:

http://webdoc.nyumc.org/nyumc/files/sun-lab/attachments/CPCB.ch02.Isolate%20cells.pdf

Here is an even easier protocol that worked for me:

http://lentiweb.com/fibroblast_isolation.php

I will say that putting the ear pieces under coverslips does help.

In most cases, these fibros won't live very long - 6-10 passages - before becoming senescent.

What are your plans for them? Have you thought of using MEFs, which grow like crazy and live a bit longer?

For tails, most protocols I found were much more involved and the ears worked fine for me...

Good Luck.

I have done this with mouse ear punches and it is pretty straightforward. I tried tails but it wasn't as easy. The fibros will actually migrate out of the ear/tail pieces and cover the dish. You basically just chop the ear with a razor blade, put it in trypsin, then plate it in media.

In a few days, you just pick out the ear pieces and all you are left with is fibros.

I will say it can take a 3-7 days so don't give up on the culture too early!

This website has a protocol from Current Protocols:

http://webdoc.nyumc.org/nyumc/files/sun-lab/attachments/CPCB.ch02.Isolate%20cells.pdf

Here is an even easier protocol that worked for me:

http://lentiweb.com/fibroblast_isolation.php

I will say that putting the ear pieces under coverslips does help.

In most cases, these fibros won't live very long - 6-10 passages - before becoming senescent.

What are your plans for them? Have you thought of using MEFs, which grow like crazy and live a bit longer?

For tails, most protocols I found were much more involved and the ears worked fine for me...

Good Luck.