[Zbrafish] Re: Zbrafish Digest, Vol 26, Issue 5

david stachura via zbrafish%40net.bio.net (by davidstachura from gmail.com)
Wed Jul 11 12:18:37 EST 2007


my suggestion would be the following:

remove the chorions however you can (just do it manually).

then, treat the embryos with liberase (Roche) according to their  
instructions.  different tissue types, thicknesses, etc. vary.  i can  
give you advice for dissociating zebrafish, but that may not help  
much.  basically, we add liberase to hanks balanced salt solution  
with calcium and magnesium.  i gently shake them on my vortexer at  
37C for 30min-hour (depends on the age).  at the end, i filter  
through .45uM mesh, and wash out the liberase well.  that should do  
the trick.

let me know if you need any further assistance.

David Stachura, Ph.D.
Cell and Developmental Biology
University of California at San Diego
9500 Gilman Dr.
Natural Sciences Building 6105
La Jolla, CA 92093-0380
davidstachura from gmail.com
dstachura from ucsd.edu
http://www.biology.ucsd.edu/labs/traver

TEL    (858) 534-6955
LAB    (858) 822-5863
FAX    (858) 822-5740


On Jul 11, 2007, at 10:00 AM, zbrafish-request from oat.bio.indiana.edu  
wrote:

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>    1. dissociated cell culture (meller from pdx.edu)
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> Date: Tue, 10 Jul 2007 11:50:04 -0700
> From: meller from pdx.edu
> Subject: [Zbrafish] dissociated cell culture
> To: bionet-organisms-zebrafish from moderators.isc.org
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> I'm a graduate student at Portland State working with Killifish.  We
> are trying to determine the best method for dissociating cells.  Any
> suggestions for what works best?  Killfish have very thick chorions so
> pronase may not be the best option.
>
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> End of Zbrafish Digest, Vol 26, Issue 5
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