The human brain governs bodily functions with exceptional complexity, yet neuroscience research is hindered by limited access to authentic human tissues, restricted availability of region-specific specimens, and a lack of physiologically relevant experimental models. Brain organoids and disease-specific brain organoids have emerged as transformative research platforms to address these bottlenecks. This review systematically summarizes advances in normal brain organoids, including cortical, cerebellar, meningeal, cerebrovascular, and blood–brain barrier models, as well as disease organoids recapitulating glioma, neurodegeneration, psychiatric disorders, neurodevelopmental defects, epilepsy, stroke and other neurological conditions. We highlight seven key translational advances of brain organoids in targeted therapy development, drug discovery and repurposing, brain–organ crosstalk, tumor brain metastasis, and longevity research. We further discuss the frontier interplay between carbon-based brain organoids and silicon-based artificial intelligence. Integrating stem cell biology, tissue engineering and clinical neuroscience, brain organoids greatly advance mechanistic research of neurological diseases and provide promising platforms for personalized medicine and regenerative therapeutics.

Glioblastoma (GBM) is a highly aggressive brain tumor characterized by rapid growth and high heterogeneity, posing challenges for fundamental research and personalized drug screening due to the lack of suitable models. GBM organoids serve as an innovative research tool, providing a valuable model for studying the biological characteristics of GBM. In this study, we successfully generated 4 GBM organoids and employed enzymatic digestion and mechanical fragmentation techniques for subsequent cultivation. Through continuous observation, pathological assessment, and RNA sequencing (RNA-seq), we observed that all the organoids generated through both methods demonstrated good growth characteristics. The organoids derived from mechanical fragmentation not only achieved a two-dimensional (2D) area of ~ 1.5 mm² but also exhibited distinct vascular structures. The organoids derived from enzymatic digestion achieved a 2D area of approximately 0.8 mm². Furthermore, RNA-seq analysis has revealed that organoids cultured using two distinct methods exhibit a heterogeneous cellular composition, comprising a total of 20 cell types (endothelial, immune cells ...). Our studies show that both methods successfully maintained the essential characteristics of GBM, encompassing its distinctive tissue structure and gene expression patterns. Each method exhibits its own attributes, contributing to the understanding of GBM organoids.

Intrahepatic cholangiocarcinoma (ICC) is a highly lethal malignancy associated with significant morbidity, necessitating the urgent development of an effective chemotherapeutic assay for ICC patients. In this study, we have successfully established an advanced culture method for ICC organoids that can be utilized with both single-cell assembly and tissue fragmentation initiation techniques. These ICC organoids maintain the morphological characteristics, including mutation profiles and frequency (46.9% in organoid and 48.5% in tumor tissue) of IDH1 genes, and 1733 high-frequent overlapped mutated genes (94.2%). Additionally, ICC biomarkers such as CK7 and CK19 also maintain a similar pattern compared with the original tissue. Furthermore, RNA-seq analysis reveals upregulation of immune-related genes in single-cell assembly organoids. The significantly changed genes including IL9R (4.4-fold), IL2RB (3.2-fold), CCR4 (3.5-fold), TESPA1 (4.4-fold), ZAP70 (4.3-fold) and CD6 (4.3-fold) in log scale. These evidence both indicating the presence of viable and active immune cells. Overall, our findings present an advanced and user-friendly culture approach for generating ICC organoids adaptable to diverse experimental objectives.

Ovarian cancer, a common gynecologic tumor, is associated with a high mortality, due to challenges in early detection within the reproductive system. According to our previous research, cultivating patient-specific organoids from mechanically sheared tissues can be utilized for drug response evaluation but has limitations for high-throughput screening efficiency due to their inconsistent size. In this research, we focused on organoids developed from single-cell suspensions to address the critical requirement for uniformity in organoid size. By the day 3 of culture, single-cell suspensions rapidly and spontaneously aggregated into spherical structures with a more consistent size. Notably, the organoids of sample OVA-37 were ten times larger after 8 days of culture. Transcriptomic analysis was used to compare the two organoid culture techniques, demonstrating that the variations between different organoid culture methods were minimal, with higher variability observed among patients. Gene set enrichment analysis (GSEA) revealed only minor discrepancies in specific pathways, such as TGF-β and tight junctions. Furthermore, treatment with carboplatin in a 96-well plate setup resulted in reproducible drug responses, as evidenced by coefficients of variation lower than 40%. This finding suggests that single-cell suspension-cultured organoids can be employed for reproducible high-throughput drug screening. This approach holds potential for personalized drug screening in ovarian cancer and may contribute to the development of novel therapeutic strategies.

Surgery is the primary treatment for skin tumors, but it can result in scarring and distress for patients. Developing alternative therapeutic methods necessitates suitable in vitro models, which are currently limited in accurately representing the in vivo cell types and microenvironment of skin tumors. Here, we present a practical approach for creating patient-derived skin tumor organoids that effectively replicate the histological characteristics and mutational profiles observed in clinical tissues. Utilizing single-cell sequencing, we identified up to 11 distinct cell types within the organoid samples, encompassing various skin system cells and immune cells. Furthermore, we demonstrate the applicability of Dermatofibrosarcoma protuberans (DFSP) organoids for assessing their responses to imatinib and metformin. Our findings reveal that metformin, in contrast to imatinib, can modulate the expression of downstream genes through immune signaling pathways. Our results underscore the ability of DFSP organoids to preserve key features of clinical tissues, including the presence of multiple cell types, especially immune cells. Importantly, our organoids provide a convenient approach for investigating the effects of drugs and elucidating underlying molecular mechanisms.