Organoids are obtained through self-organization of cells that grow into miniaturized equivalents of organs comprising their micro-anatomical structure. Cerebral organoids (COs) are normally formed through consecutive steps including embryoid body (EB) formation, neuroepithelial induction, Matrigel embedment, and further maturation. However, Matrigel represents a major obstacle for the transition of organoid technology to clinical applications due to its tumorigenic potential, batch-to-batch variation, and poorly defined content. There is, therefore, a major need for alternative matrices to grow biologically safe organoids. Self-assembling matrices based on peptide amphiphiles (PAs) are an attractive alternative given their well-defined chemistry, engineered bioactivity, and nanofibrous architecture. Here, we offer a co-assembling platform for the culture and induction of COs, that integrates positively charged and laminin-mimetic PAs with the negatively charged hyaluronic acid (HA) functionalized with tyramine (HA-Tyr). The developed chemically defined platform enables tuneability of stiffness to investigate its effect on CO induction and growth. Multi-omics approaches including transcriptomics, proteomics, and metabolomics demonstrated the ability of HA-Tyr/PA gels to induce EBs towards COs displaying structures similar to those obtained by the conventional Matrigel-based method. EBs have been shown to more likely be induced towards COs and display a similar biomolecular structure in soft matrix compared to stiff matrix, confirmed by the multi-omics analyses. The developed material is expected to serve as a safe and practical matrix for the induction of COs, which might accelerate the transition phase of COs from laboratory to the clinics.