A total of 22 teams will contest the league, including 15 sides from the 2023–24 season, three relegated from the 2023–24 La Liga, and four promoted from the 2023–24 Primera Federación.
On 26 May 2024, Real Valladolid became the first side to mathematically be promoted, assuring an immediate return to the top flight following 3–2 win against Villarreal B.
The first team to be relegated from La Liga were Almería, after a 3–1 loss to Getafe on 27 April 2024, ending their two years stay in the top tier.
The third and final team relegated to Segunda was Cádiz, after a 0–0 draw against Las Palmas on 19 May 2024, ending their four-year stay in top tier.
The second team to earn promotion was Deportivo La Coruña following a win against Barcelona Atlètic on 12 May 2024, ending a four-season stint at the third division.