Lab-On-a-Brane (LOB) represents a class of Lab-On-a-Chip (LOC) integrating flexible, highly gas permeable and biocompatible thin membranes (TMs). Here we demonstrate the potentiality of LOBs as cell biochips promoting 3D cell growth. The human cancer cells MCF-7 were cultured into standard multiwells (MWs) and into polydimethylsiloxane (PDMS) MWs, LOCs, and LOBs of different wettability. Surface treatments based on oxygen plasma and coating deposition have been performed to produce hydrophilic, hydrophobic, and oleophobic chips. By a comparison between all these chips, we observed that 3D cell aggregation is favored in LOBs, independent of substrate wettability. This may be attributed to the TM flexibility and the high oxygen/carbon dioxide permeability. Ultimately, LOBs seem to combine the advantages of LOCs as multi-well microfluidic chips to reduce operation time for cell seeding and medium refresh, with the mechanical/morphological properties of PDMS TMs. This is convenient in the perspective of applying mechanical stimuli and monitoring cell stiffness, or studying the metabolism of molecules permeable to PDMS membrane in response to external stimuli with interesting outcomes in cellular biology.