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Once you've unlocked everything, I almost always go to the Cathedral because it's more fun later.
Which path you should take depends on if you took the polaroid (go up) or the negative (go down) after defeating Mom. Taking the correct path will get you to the Chest level to fight ??? (if you took hte polaroid and went up) or the Dark Room level to fight the Lamb (if you took the negative and went down). Defeating both of these final bosses with every character on hard mode is required to 100% the game.
If you haven't unlocked th epoloroid/negative yet, then you need to defeat Isaac/Satan to advance that goal. For example, defeating Isaac five times will unlock the polaroid.
(Aside: Polaroid photos like the one depicted in the game don't have film negatives... that always kind of bothered me)