| Atlanta United II | |
| Austin II | |
| Bethlehem Steel | |
| Chicago Fire II | |
| Colorado Rapids II | |
| Columbus Crew II | |
| Crown Legacy | |
| FC Cincinnati II | |
| Fort Lauderdale CF | |
| Houston Dynamo FC II | |
| Huntsville City | |
| LA Galaxy II | |
| Los Angeles II | |
| Minnesota United II | |
| New England II | |
| New York City II | |
| New York RB II | |
| North Texas | |
| Orlando City II | |
| Portland Timbers II | |
| Real Monarchs | |
| San Jose Earthquakes II | |
| St. Louis City II | |
| Swope Park Rangers | |
| Tacoma Defiance | |
| Toronto II | |
| Whitecaps II | |