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Heavy spoilers of course, but you asked.
Step 1: the basics.
- Kill Mom to unlock the Womb floors.
- Kill the Heart (Womb floor boss) 10 times to unlock Hush (accessible on further runs by killing the Heart under 30 minutes).
Step 2: preparing other requirements:
- Kill the Heart an 11th time to unlock a permanent access to the Cathedral and Sheol.
- Beat the Cathedral or Sheol floor 5 times to unlock a specific item (the Polaroid and the Negative, respectively). Only one is needed.
Step 3: requirements, second half.
- Kill Hush 3 times to unlock the alternative path. Its starting point is at the end of Basement 1, a door that requires a key. Further doors to stay on the path will require two bombs and two hearts of damage (unless you're the Lost).
- On that path, complete the knife (its pieces are somewhere on Downpour 2 and Mines 2), use the knife on the flesh door after Mom's fight on Mausoleum 2, kill the boss behind it, beat the Corpse floor. This will unlock a new path (the Ascent) accessible from the Depths 2 floor.
Step 4: now you're ready, here's how Tainted characters are unlocked.
- To open that door on Depths 2, kill Mom, take the Polaroid or Negative, teleport out of the boss room (if you have nothing to teleport, a marked skull somewhere on the floor is guaranteed to drop a Fool card when bombed).
- Touch the door, you'll lose your Polaroid/Negative. Go on that path, Home is at its end. The first time you go there you're guaranteed to get the Red Key, so you can unlock a Tainted character immediately.
- On further runs, the Red Key isn't guaranteed to show up anymore. But it's not needed: drop a trinket in a treasure or boss room during the run. Then, during the Ascent, it will be transformed into a one-use version of the Red Key (so you can unlock other Tainted characters without the actual Red Key).
Doing this process as Isaac will unlock Tainted Isaac. Repeat for each character.
Note that this doesn't even apply to the first character you do the process with.
For part 3, the knife pieces and the alt floors, there's some luck involved with getting the key to open downpour, and the two bombs to open the door to the mines. It's rare that you aren't able to though, since you only need to go to downpour 2 and mines 2 to get the knife pieces, meaning you can delay going there by a floor if you need to. It's more often that I lock myself out of mines because I forget to save the bombs you need to get there, than the game just not giving me enough.