I'd say there's a few big questions you need to answer to help make this decision. The biggest of which is, what does your site/product/customers do? If you're managing an application (like say a CMS or shopping cart) then you need to understand that many hosts will not be upgrading for a while because they wait for officially supported packages/RPMs for the OS they use, and they take time to build, test and release.
If this is just a custom site running on a dedicated server the same question can be some what important. While, in this case, you can always compile by hand that may not necessarily be the best idea if you're as anal about a clean, organized, server as I am. Like Jason mentioned, there are supposed to be significant speed improvements, and as WishCow said, if you use create_function() you can now ditch them for cleaner code.
Also keep in mind any third party libraries/extensions (PECL, PEAR, Zend Framework, Drupal, Wordpress, or custom written) that you're using that may not work on 5.3 yet for any given reason.
A dry-run upgrade is always better if it's possible. I have a server sitting next to my desk at home that I can use to upgrade and turn on a profiler to see what the performance of my site(s) looks like and make sure everything works. If you can't do this then you have to be careful because there's a chance you'll have some down time if things don't go smoothly.