The reigning European champions will welcome the WSL title holders to Estadi Olimpic Lluis Companys for Saturday's first leg.
Match preview
Having already clinched the Spanish Super Cup, Barcelona are hoping to win three more pieces of silverware before manager Jonatan Giraldez leaves in the summer to become the new head coach of NWSL side Washington Spirit.
The Catalan giants are 12 points clear at the top of the Liga F table with seven matches left to play, while they are set to face Real Sociedad in next month's Copa de la Reina final.
They are also looking to retain their European crown after sealing their place in the semi-final with a 5-2 aggregate win over Brann in the last eight.
The two-time European champions are now targeting a repeat of last season's 2-1 semi-final victory against Chelsea.
Barcelona have not lost a two-legged Champions League tie since they were knocked out by record winners Lyon in the 2017-18 quarter-finals, highlighting the size of the task that awaits Chelsea over the next two matches.
Like Barcelona, Chelsea will also bid farewell to their manager at the end of the season, with Emma Hayes set to become the new head coach of the USA Women's national side.
Hayes's hopes of winning a quadruple or a treble have been quashed by a League Cup final defeat to Arsenal and an FA Cup semi-final loss against Manchester United.
The Blues bounced back from those disappointing results to record a 3-0 victory in Wednesday's home league meeting with 10-player Aston Villa thanks to goals from Aggie Beever-Jones, Maika Hamano and Kadiesha Buchanan.
After moving top of the WSL table on goal difference, Chelsea will now turn their attention to European action after booking their place in the semi-final with a 4-1 aggregate victory in last month's quarter-final tie against Ajax.
The Blues, who are dreaming of winning their first European title, are hoping to reach the Champions League final for the first time since 2020-21 when they ultimately finished runners-up to Barcelona.
Barcelona Women Women's Champions League form:
W
W
W
D
W
W
Barcelona Women form (all competitions):
Chelsea Women Women's Champions League form:
D
W
W
W
W
D
Chelsea Women form (all competitions):
Team News
Barcelona are still unable to call upon the services of Mapi Leon, who has not played since November due to a knee injury.
Cata Coll is set to replace Sandra Panos in goal, while Irene Paredes, Ingrid Engen and Ona Batlle are expected to come into the backline after being left on the bench at the weekend.
Keira Walsh, Caroline Graham Hansen and Mariona Caldentey are also set to return to Barcelona's starting lineup for the crucial Champions League fixture.
As for the visitors, they are expected to be without Millie Bright, Nathalie Bjorn, Aniek Nouwen, Maren Mjelde, Mia Fishel and Sam Kerr.
After rotating her side in midweek, Hayes is set to recall a number of players, including Eve Perisset, Erin Cuthbert and Melanie Leupolz.
Johanna Rytting Kaneryd, Lauren James and Mayra Ramirez are also poised to return to the starting lineup for Saturday's first leg.
Barcelona Women possible starting lineup:
Coll; Bronze, Paredes, Engen, Battle; Bonmati, Walsh, Guijarro; Graham Hansen, Caldentey, Paralluelo
Chelsea Women possible starting lineup:
Hampton; Carter, Buchanan, Charles; Leupolz, Cuthbert; Rytting Kaneryd, James, Reiten; Ramirez