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The club was started in the fall of 2009 and co-owned by Erik Tammer of Tammer Sportmanagement and Mike Mossel of Business and Sports Performance. Tammer is a former professional soccer player in the Netherlands who played for clubs such as Heerenveen and Sparta Rotterdam, while Mossel played in Europe for RBC Roosendaal and Turnhout, and in the USL for the Cincinnati Riverhawks following his graduation from Xavier University.

The team played its first competitive game on May 8, 2010, a 3–3 tie with the Cincinnati Kings. The first goal in franchise history was scored by Eddie Hertsenberg.

On July 8, 2010, the Lions announced that they would be self-promoting to the USL Second Division for the 2011 season, as well as adding a brand new women's team in the USL W-League. During the announcement team owner Mike Mossel said "The promotion to USL-2 and development of a W-League club was just a matter of time. To be the best, we must begin to play among the upper-echelon clubs. We want to set a standard, and I believe we have done that so far in the PDL in our first season. But more importantly, our decision was made with our Premier Academy in mind.". With the merger of the USL First and Second Divisions to form USL Pro in 2011, the Dutch Lions were announced as a founding team.

In 2012, the Lions reached the Quarterfinals of the 2012 Lamar Hunt U.S. Open Cup, where they defeated the MLS Columbus Crew 2–1 in the third round, then defeated the Michigan Bucks in the fourth round in extra time. They were knocked out of the tournament in the quarterfinals by eventual champions Sporting Kansas City 3–0 in front of an away record 15,167 fans.

In 2013, the Lions reached the USL Pro Playoffs for the first time with a record of 10 Wins, 7 Ties and 9 Losses. They lost to the Richmond Kickers 1–0 in the quarter-finals. Gibson Bardsley received Second Team All-League Honors with 9 goals and 7 assists.

In 2014, the Lions partnered with West Carrollton City Schools and Dr. Suresh Gupta of the Dayton Outpatient Center to install a new playing surface in the West Carrollton High School Stadium as part of a 30-year deal. The original stadium was renamed to the Dayton Outpatient Center Stadium in recognition of the significant investment made by Dayton Outpatient Center.

Renovations started on March 10, 2014, and were completed with a ribbon-cutting ceremony on May 9, 2014. The Motz Group, a Cincinnati-based company that specializes in building playing fields, replaced the existing grass field with artificial turf at a cost of $450,000 to $475,000. The combined costs for the renovations to the stadium and installation of lights on practice fields are estimated at $529,000.

The opening match at DOC Stadium, with the presence of 2-Time FIFA World Player of the Year Ruud Gullit, was a United Soccer League professional soccer game on May 10, 2014, that saw the Dayton Dutch Lions tie the LA Galaxy II, 1–1. A home record attendance of 1,527 was announced.

In 2018, the Lions had a record-setting season, matching its best record ever, winning the Great Lakes Division championship and earning qualification for the 2019 U.S. Open Cup.