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Darkspore gameplay was similar that of other action-RPG titles, such as Torchlight and Diablo, with a top-down third-person viewpoint, 'click-to-attack' and 'click-to-move' controls, and special abilities activated by using keys 1-5.

Darkspore had high-rendered modern graphics and textures, and visceral gameplay that moved at a slightly calmer pace than most other 'hack and slash' games to aid its strategic edge.

The player would have been able to collect up to 100 heroes, made up of 25 heroes each with 4 variants. Each variant is assigned a letter of the Greek alphabet in order of when they're unlocked (Alpha, Beta, Gamma, Delta) which were of the same type and class and wielded the same weapons, but had their own unique look, different stats, and in addition to the 3 abilities which were universal across every variant of the same hero, each of the 4 variants had one separate ability of their own, called 'Variant Abilities'.

Each player could choose a squad deck of 3 heroes to take into every mission, with up-to 3 squads. The right selection and combination of heroes was vital, as their abilities should compliment each others' combat effectiveness, and their types and classes should be selected so as to counter resistances according to the enemies being faced. The player could only control one of the deck at a time, though was able to swap freely between the three. Switching was useful for protecting heroes that were low on health or energy until they could be healed or re-energized, also switching knocks-back enemies and cleanses debuffs. Every hero had its own 'squad ability', an ability which could be used by any of the heroes in the player's squad-deck at any time without switching.

The game's three primary pillars of gameplay were Collection, Customization and Combat. Players would have to rise to the challenge to collect new heroes and many special items, customize and improve those heroes in the editor using the collected items, and then take their selection of the best heroes for the job planet-side to engage in strategic combat and reap the spoils of victory.

Darkspore boasted great replayability, and was designed to have much worth in being played over again several times. The game featured a procedural AI Director that dynamically controlled enemies, and an assessment system that encouraged players to chain multiple missions and earn better loot, a 'Risk versus Reward' system that gave players the option to push their luck rather than cash-out in a risky but exciting gamble. However, if the player chose to push their luck but fails the following mission, all would be lost. Darkspore left plenty of room for experimentation.

The game emphasized on its co-op elements, as the best campaign experience came from playing as a team with a friend or two.

Darkspore also featured an online Player vs. Player feature, (commonly known as PvP), a component that allowed players to pit their customized heroes that they have collected in normal gameplay, against other players' heroes, in a multiplayer battle arena. There would be up to 3 rounds of PvP. Like in the campaign, the player would have a deck of 3 heroes each that the player could swap control of using a universal ability to balance their health, energy, and abilities. The game supported 1 vs. 1 and 2 vs. 2 battles, letting up to four players at a time to join a match.

PvP battles were much more challenging and tactical than that of certain other similar games, as defeating an opponent required players to use all manner of heroes, abilities, and techniques to eliminate all three of the opponent's heroes, which were likely just as powerful. Online PvP featured online matchmaking, ranking, and leaderboards.

Darkspore's co-operative multiplayer campaign was the main focus of the game. Up to four players, each with their own squad deck of 3 customizable heroes, could join in on a campaign game, battling enemies together, forming team tactics, and trading loot.

However, there was one extra challenge in Co-op gameplay, Darkspore Operatives, special enemies that trapped a member of your team, as well as draining their health.

There were 5 different Darkspore Agents, each of a different genesis. They were:

  • Cyber - Omicron - Locked down players in a cage whilst healing itself with a repair drone.
  • Bio - Vaulting Amphiod - Could not be interrupted until dead. They could lock-down two players at once for a short time.
  • Plasma - Magmatic Brute - Summoned two minions with 1 hp, Volt, who could lock down a hero and Cinder, who could ignite heroes. If these minions died, the Magmatic Brute would resummon them. The Magmatic Brute could punch pets while its minions were attacking a hero.
  • Necro - Haunt Strider - Emerged from stealth and snared a hero, and upon getting closer would lock them down.
  • Quantum - Gravitic Confiner - Was able to lock down heroes at a very long range. Would teleport away when low on hp, appearing shortly after again locking down another hero. Stunning the confiner when it was low on hp would prevent it from teleporting until either dead or the stun wore off.

The only way to free that member was to kill the Darkspore Agent or put them to a affect where they don't attack or push them back , putting them in the hands of their fellow teammates.

If there was only one player left, Darkspore Agents would be unable to lock them down.