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Kill Satan 5 times to unlock the Negative. Then take it in a run, kill Satan again, and you'll go to the Dark Room.
Beating either of these floors will unlock the Mega Satan fight (located on the Chest and Dark Room floors, though you'll have to get the key to its door during the run).
If you have Repentance, there's also Mother and the Ascent path. Kill Hush 3 times to unlock Mother's path (killing it will unlock the Ascent on further runs).
You didn't mention the Boss Rush, so there's that too (kill Mom under 20 minutes). And Greed(ier) mode of course.
Reminder: to get all unlocks, all these things must be done as all characters.
also the worst fight