This is a cached page for the URL (http://www.scienceboard.net/resources/protocols.asp?action=article&protocol_id=624&criteria=). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
The Science Advisory Board - Protocols, Product Reviews, Member Forum, and Science News
Note: You are seeing this message either because your browser has not loaded our stylesheets, or because your browser does not support stylesheets (CSS). Please upgrade to a relatively modern browser to improve your experience. Not sure what to upgrade to? Try Firefox.
The Science Advisory Board
Screen Name: 
 
Password: 
 

Protocols

Welcome to The Science Advisory Board's extensive database of research protocols for the biomedical sciences. Protocols are organized by techniques and are fully searchable by key word(s). Protocols are divided into a brief description of the methods, a step-by-step overview of the procedure, a collection of recipes, the supplies needed, and any helpful tips. Members submitting protocols will be rewarded through the ViewPoints system.
... back to articles list

Endothelial wound healing (cell migration) assay

Posted on Friday, January 26, 2007

Description
This is a simple assay that can be used in any cell culture lab setup to test the effect of different compounds on endothelial cell migration.

Procedure
Grow endothelial cells in complete media.

Day 1: Trypsinize cells and add 4 x105 cells per well in a six well plate in complete media. After 8 h, change media to starvation media. Starve cells overnight in this media.

Day 2:
1) Remove media from cells.
2) Make a scratch wound across each well of the 6-well plate using a pipette tip. Make sure to use a very fine tip so that the cells come off the plate and the cell free area has sharp clear edges.
3) Wash three times with starvation media to remove any loosely held cells. Make sure that the wound area is free of cells.
4) Add starvation media containing required concentration of compound to be tested in 3 ml volume per well.
5) Take 0 hr. images.
6) Incubate at 37 degree C for 6 h. Take images.
7) Compare 0 and 6 hr images and calculate area of the wound closed using image J software.


Recipes


Supplies
Cell culture media
FBS (Invitrogen)
Complete media: Medium with FBS supplemented to 10%
Starvation media (SM): Medium with no FBS
Endothelial cells such as HUVEC, BAEC

75 cm2 vented flask (gelatin coated) (Biocoat)
gelatin coated 6-well plate (Biocoat)


Tips
1) The media used is the specific media for that cell type.
2) One can also try using media containing low percentage FBS to obtain faster closing of the wound.
3) The time of incubation can be increased or decreased based on the type of cells used (how fast they migrate) and the compound being tested.


Submitted by: sci2k1

Scientific & Medical
Experts Needed!

The Science Advisory Board is the world's most established network of life scientists!

Voice your opinions on companies, products, protocols and even humor in a lively, real-time, interactive Online Community of over 42,000 life science & medical professionals.

Redeem generous rewards for participation in studies, contributing website content and referring colleagues.

Join Right Now!
(It's Free!)

Search This Site
only search scienceboard.net
only search Forums
What's this?