| 08 Oct 21 | r16        | New England Revolution IINew England R | 1 - 0 | Toronto FC IIToronto FC II |
| 09 Oct 21 | r16        | North Carolina FCNorth Carolin | 2 - 3 | Chattanooga Red Wolves SCChattanooga R |
| 11 Oct 21 | r16        | North Texas SCNorth Texas S | 1 - 2 | Richmond KickersRichmond Kick |
| 11 Oct 21 | r16        | Fort Lauderdale CFFort Lauderda | 4 - 4 | Tormenta FCTormenta FC |
| 14 Oct 21 | r16        | FC TucsonFC Tucson | 0 - 2 | North Carolina FCNorth Carolin |
| 16 Oct 21 | r16        | Richmond KickersRichmond Kick | 3 - 2 | Forward Madison FCForward Madis |
| 17 Oct 21 | r16        | Chattanooga Red Wolves SCChattanooga R | 1 - 2 | Toronto FC IIToronto FC II |
| 17 Oct 21 | r16        | Greenville Triumph SCGreenville Tr | 1 - 0 | New England Revolution IINew England R |
| 17 Oct 21 | r16        | Union OmahaUnion Omaha | 4 - 1 | North Carolina FCNorth Carolin |
| 17 Oct 21 | r16        | FC TucsonFC Tucson | 3 - 1 | Tormenta FCTormenta FC |