The criteria for the list includes academic reputation, peer assessment, graduation rates, retention rates, faculty resources, financial resources and alumni giving.
Creighton edged out Butler University in Indianapolis and Drake University in Des Moines for the top spot.
Its the 15th consecutive year Creighton has been No. 1 in the category.
"Fifteen consecutive years as the top-ranked University in the Midwest is an immense accomplishment for Creighton University and it speaks volumes about the caliber of students, alumni, faculty, staff and programs," Creighton president Rev. Daniel Hendrickson said. "We continue to be grateful and inspired that Creighton is recognized for the academic, global and spiritual engagement to our community."