Atleti generally play at Estadio Cerro del Espino, a 3,376-seater ground which is shared with Segunda Division side CF Rayo Majadahonda, but were given the opportunity to use the Wanda Metropolitano for their top-of-the-table clash with Barca.
The two teams were just six points apart heading into the Primera División Femenina match up, with Lluis Cortes's visitors chasing their table-topping hosts.
That point gap was reduced to three in front of the record crowd, however, with Asisat Oshoala netting the opener to take her tally to five goals in five games, while England international Toni Duggan took mere minutes to add a second after her late introduction from the bench.
The 2-0 result will no doubt feel like revenge of sorts for the Catalan side, having been ousted from the cup by Atleti in mid-February – especially given the enormous turnout in the Spanish capital.
The previous record attendance for a women's club match stood at 48,121 – a Copa de la Reina clash in which Athletic Bilbao hosted Atleti, with the latter winning 2-0 thanks to goals from Angela Sosa and Aurelie Kaci back in January.
In women's football as a whole, the 1999 World Cup final still holds the overall record, with 90,185 fans turning out at the Rose Bowl in California to see the United States Women's National Team triumph over China in a penalty shoot-out after the game had been tied at 0-0 following extra time.
Atleti, meanwhile, must dust themselves off ahead of a trip to Levante and restore momentum as they look keep their Primera Division lead intact and build confidence before their Copa final clash with Real Sociedad in May.