Molecular and Proteomic Profiling of Human Cerebral Organoid‑Derived Small Extracellular Vesicles

During the culture of organoids and spheroids, the generated tissues are capable of secretion of small Extracellular vesicles (sEVs) into their microenvironment which are functional bioactive molecules. With the help of the advancements such as easy to control increase culture condition homogeneity and reproducibility and high level of control over the microenvironment in the on-chip platform development field, we differentiated iPSC derived COs under three different culture conditions as static, by utilizing orbital shaker and our novel dynamic Organoid-on-a-Chip platform during 30 days of cerebral organoid maturation step to compare the different growth condition effects on developmental stages and EV profiles, we investigated the differences of organoid features in terms of bulk RNA analysis, protein expression and EVs characterizations.

Principal component analysis (PCA) of organoid transcriptomes indicate strong culture condition-specific modulation of gene expressions of the organoids. Proteomic results from EVs showed similarities between culture conditions in the early stage, it was observed that differences in conditions in the late stage affected the proteome in terms of cellular maintenance, stress response, and tissue-specific maturation. sEVs treatment showed significant decrease in viability as well as migration of the U87 cells especially orbital shaker condition.