Some demonstrative embodiments include devices, systems and methods of multi-user uplink transmission. For example, an apparatus may include a transmitter to transmit a multi-user (MU) downlink transmission to a plurality of wireless stations; a receiver to receive from the plurality of wireless stations a plurality of acknowledgement (ACK) frames, at least one ACK frame from at least one wireless station including an uplink scheduling request indicating uplink resources requested by the wireless station; and a scheduler to schedule an uplink transmission from the at least one wireless station based on the uplink scheduling request, the transmitter to transmit at least one scheduling frame including scheduling information of the scheduled uplink transmission.