We propose and demonstrate vertically oriented self-assembly of colloidal quantum wells (CQWs) that allows for stacking CdSe/CdZnS core/shell CQWs in films for the purposes of amplified spontaneous emission (ASE) and random lasing. Here, a monolayer of such CQW stacks is obtained via liquid-air interface self-assembly (LAISA) in a binary subphase by controlling the hydrophilicity/lipophilicity balance (HLB), a critical factor for maintaining the orientation of CQWs during their self-assembly. Ethylene glycol, as a hydrophilic subphase, orients the coalition of these CQWs into self-assembled multi-layers in the vertical direction. Stacking CQWs into large micron-sized areas as a monolayer is facilitated by adjusting HLB with diethylene glycol addition as a more lyophilic subphase during LAISA. ASE was observed from the resulting multi-layered CQW stacks prepared via sequential deposition onto the substrate by applying the Langmuir-Schaefer transfer method. Random lasing was achieved from a single self-assembled monolayer of the vertically oriented CQWs. Here, highly rough surfaces resulting from the non-close packing nature of the CQW stack films cause strongly thickness-dependent behavior. We observed that in general a higher roughness-to-thickness ratio of the CQW stack films (e.g., thinner films that are intrinsically rough enough) leads to random lasing, while it is possible to observe ASE only in thick enough films even if their roughness is relatively higher. These findings indicate that the proposed bottom-up technique can be used to construct thickness-tunable, three-dimensional CQW superstructures for fast, low-cost, and large-area fabrication.