1

The greatest medical ninja should have learned it somewhere.

Tsunade's skills extend beyond the typical medical ninjutsu. She earned fame during the Second Shinobi World War for being the only person able to make antidotes for the poisons of Sunagakure's Chiyo. Similarly, she can make sleeping potions that are undetectable to even the most skilled ninja. She is knowledgeable enough to perform autopsies and examine damage on a cellular level.

Source from Narutopedia.

5

There's no background information explaining how Tsunade became a medical-nin or how she learned medical ninjutsu. She didn't have a medical specialist mentor. Maybe she did have one, but there's no source or background information to prove it.

There is a possibility that she learned medical ninjutsu all by herself, as she often read other clans' medical books to solve particular problems.

1

As others have answered that there is no concrete evidence on who taught her medical ninjutsu but we can theorize.

My guess would be the First Hokage. Consider the following information

  1. He doted on his granddaughter and spent time with her to teach her gambling. (He'd said he shouldn't have taught her gambling)
  2. He was the greatest ninja of his time. He could self "heal his wounds without weaving any hand signs"

Logically I'd guess Tsunade was influenced by the first and may have even got the basics of medical ninjutsu from the first Hokage. She then gained tremendous experience in the Ninja Wars and doing missions with the Sannin. The real life experience with the large amount of chakra and aptitude made her the greatest living medical-Nin in Narutoverse. She no doubt build on the initail knowledge travelling and reading about in other books, like Nara Clans Herbology.

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