If I were to putting the starter, I will put 20 starting character, 24 unlockables character, and 4 update character

Starters

1. Pico (Pico's School)
2. Nene (Pico's School)
3. Darnell (Pico's School)
4. Girlfriend (Friday Night Funkin)
5. Son (Dad n Me)
6. Alien (Alien Hominid)
7. Hank (Madness Combat)
8. John (Tankmen)
9. P-Bot
10. Alloy (Alloy Arena)
11. Salad Fingers
12. Henry Stickmin
13. Edd (Eddsworld)
14. Meat Boy
15. Isaac (The Binding Of Isaac)
16. Phil (Riddle School)
17. Ena
18. Doki and Nami (There She Is)
19. Cassette Girl
20. Polandball

Unlockable Characters

1. Cassandra (Pico's School)
2. Convict (Pico's School)
3. Piconjo
4. Boyfriend
5. Knight (Castle Crasher)
6. Hatty (Battleblock Theater)
7. Grunt (Madness Combat)
8. Tricky (Madness Combat)
9. Fancy Pants
10. Samurai Asshole
11. Smash Girl (Powertrip)
12. Blockhead
13. Bitey (Brackenwood)
14. Stan (Nightmare Cops)
15. Satina
16. Strawberry Clock
17. Larry (Larry of Karryon)
18. Skid and Pump (Spooky Month)
19. Ami (Gwain Saga)
20. Gildedguy
21. J.Y. (JYGame)
22. Flippy (Happy Tree Friends)
23. Homestar (Homestar Runner)
24. Firey (Battle For Dream Island)

Update

1. Otis (Pico's Cousin)
2. Doobus Goobus
3. Peppino (Pizza Tower)
4. Cloog (Major Cloog)

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Green = Playable

Red = Unlockable

Yellow = Update