The visitors hold a two-goal lead from last week's first leg in Santiago and it is now up to the home side to come up with a much-needed response.
Match preview
Heavily fancied to get the job done on home turf, Colo Colo lived up to expectations courtesy of a 2-0 victory against Internacional at the Estadio Monumental David Arellano on Wednesday.
Gustavo Quinteros's men took the lead in the first half through Juan-Martin Lucero before Pablo Solari sealed the deal 10 minutes into the second period.
Los Albos played out a thrilling 1-1 draw against Everton in their subsequent league outing on Friday and they are now unbeaten in five games across all competitions.
However, Wednesday's visitors cannot afford to take their foot off the gas in their next involvement having suffered a heavy 4-0 defeat in their last continental affair on the road.
Internacional failed to win away from home for the second game running following a 1-1 stalemate versus Ceara in Saturday's encounter at the Castelao.
Mano Menezes's men trailed in the 20th minute to Victor Lima's penalty but Moises restored parity in Fortaleza just four minutes later.
Clube do Povo now return to their own patch where they claimed a convincing 3-0 victory against Coritiba last time out, stretching their unbeaten streak to nine games.
Wednesday's hosts set out to put up another dominant display in front of their fans and book a spot in the last eight of this year's Copa Sudamericana.
Internacional Copa Sudamericana form:
Internacional form (all competitions):
Colo-Colo Copa Sudamericana form:
Colo-Colo form (all competitions):
Team News
Rodrigo Dourado is not expected to feature as he remains sidelined with a knee problem and the 28-year-old will be a huge miss for Internacional.
Twenty-nine-year-old Rene was subbed off in the first half of his side's defeat to Colo Colo, making him a doubt for Wednesday's game.
Pedro Henrique and Alexandre Alemao are expected to return to the starting XI here after they both sat out the hosts' last outing.
Gabriel Costa was on fire in the first leg where he set up both of his side's two goals and the Peruvian will be looking to deliver the goods once again.
Esteban Pavez and Jeyson Rojas were both sent off for the visitors against Everton but the duo are eligible to play a part in this one.
Internacional possible starting lineup:
De Sousa; Moises, Mercado, Vitao, Heitor; Henrique, Gabriel, Johnny, Edenilson; Patrick, Alemao
Colo-Colo possible starting lineup:
Cortes; Lara, Falcon, Zaldivia, Suazo; Fuentes, Pavez; Solari, Gil, Costa; Lucero