Journal: bioRxiv

Article Title: Multi-omics and spatial analysis of microgravity-grown glioblastoma organoids reveals superior modeling of advanced disease after long-term spaceflight

doi: 10.64898/2026.03.06.710192

Figure Lengend Snippet: ( A ) Organoids were created with three distinct cellular compositions – GBM cells alone, GBM+monocytes, and GBM+macrophages in 1:1 mixtures. GBM+macrophage organoids were further treated with cytokines to polarize the macrophages into an M1-like or M2-like phenotype. ( B ) Overview of the hanging drop method of organoid formation (left) and schematic of organoids either free-floating or embedded in agarose in cryovials (right). ( C ) Timeline of the spaceflight experiment from organoid formation to endpoint processing. ( D ) Overview of post-flight multi-omics analysis methods of the organoids.

Article Snippet: After 48 hours, organoids were transferred from the hanging drops into cryovials (Thermo Scientific, 374088), to be grown either freely suspended in medium or embedded in agarose.

Techniques: Biomarker Discovery

Journal: bioRxiv

Article Title: Multi-omics and spatial analysis of microgravity-grown glioblastoma organoids reveals superior modeling of advanced disease after long-term spaceflight

doi: 10.64898/2026.03.06.710192

Figure Lengend Snippet: ( A ) Phase-contrast images of GBM alone, GBM+monocyte (Mono), and GBM+macrophage (MΦ) organoids after 48 hours in hanging drops. Scale bars represent 200 µm. ( B ) Images showing monocytes and macrophages (magenta) incorporated with GBM cells in live organoids (green). Scale bars represent 200 µm. ( C ) Viability staining showing live cells (green) and dead cells (red) in organoids after 47 days in sealed cryovials. Scale bars represent 200 µm. ( D ) Brightfield images of ground control and post-flight microgravity organoids after fixation and removal from agarose. Scale bars represent 200 µm. ( E ) Quantification of compactness, solidity, and form factor from the brightfield organoid images. Statistical comparisons were made using unpaired t tests (* p < 0.05, ** p < 0.01). Error bars indicate standard error of the mean (SEM). ( F ) Hematoxylin and eosin staining of 6 µm sections of ground control and microgravity organoids. Scale bars represent 200 µm.

Article Snippet: After 48 hours, organoids were transferred from the hanging drops into cryovials (Thermo Scientific, 374088), to be grown either freely suspended in medium or embedded in agarose.

Techniques: Staining, Control

Journal: bioRxiv

Article Title: Multi-omics and spatial analysis of microgravity-grown glioblastoma organoids reveals superior modeling of advanced disease after long-term spaceflight

doi: 10.64898/2026.03.06.710192

Figure Lengend Snippet: ( A ) Image of a cryovial with no successful organoid formation with the original pink media color (left) and a cryovial containing a 45-day old organoid with acidified, yellow media (right). ( B ) Images of ground control and microgravity cryovials initially loaded with organoids made with 50 cells, 100 cells, and 250 cells. Vials on the left of each image (50-cell organoids) were more pink, indicating failure to support a successful organoid, and vials on the right (250-cell organoids) were more likely to be yellow, indicating organoid growth and media consumption. ( C ) Fraction of cryovials in each condition found to contain a successful organoid at the 45-day experiment endpoint. Error bars represent SEM. Significance was calculated with Welch’s t test, * p < 0.05.

Article Snippet: After 48 hours, organoids were transferred from the hanging drops into cryovials (Thermo Scientific, 374088), to be grown either freely suspended in medium or embedded in agarose.

Techniques: Control