Journal: Advanced Healthcare Materials

Article Title: Towards Automation in 3D Cell Culture: Selective and Gentle High‐Throughput Handling of Spheroids and Organoids via Novel Pick‐Flow‐Drop Principle

doi: 10.1002/adhm.202303350

Figure Lengend Snippet: Cytocompatibility of PFD. a) Exemplary image of MCF7 spheroids embedded in basement membrane extract (BME) with PFD and by manual pipetting one hour after plating. Scale bar: 200 µm. b) Exemplary image of CRC organoids embedded in BME with PFD and by manual pipetting one hour after plating. Scale bar: 200 µm. c) Cell viability 4 h after plating either via PFD or via manual pipetting (MP). Three technical replicates, n ≥ 60 for each sample type and each condition. d) Measured luminescence signal due to Annexin V binding to exposed phosphatidylserine on the cell surface as an indicator of apoptosis 24 h post plating. Three technical replicates, n ≥ 60 for each sample type and each condition. e) Measured loss of membrane integrity as indicator of secondary necrosis 24 h post plating. Three technical replicates, n≥60 for each sample type and each condition. f) Normalized area of unprocessed cell aggregates and aggregates that were processed with the spheroid and organoid processing platform. Each condition shows data of n ≥ 60. Aggregates with diameters larger than 240 µm were not considered for aspiration and are therefore excluded from the data set. g) Exemplary proliferation of an MCF7 spheroid that was automatically embedded in BME over the course of several days. Day 1 was the day of embedding. Scale bar: 100 µm. h) Fold change in diameter of MCF7 spheroids after they were embedded in basal membrane extract with the platform. n = 34. Spheroids were automatically delivered into the hydrogel on day 1. Data was tested for normal distribution and the two‐tailed t ‐test was performed. P‐ values ≥ 0.05 were considered nonsignificant (n.s.).

Article Snippet: Briefly, organoids were cultivated in domes of 4 mg mL −1 basement membrane extract BME (Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2, Pathclear, R&D Systems, Inc., USA).

Techniques: Membrane, Binding Assay, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Hippo pathway suppression reprograms TNFα-primed glioblastoma extracellular vesicles transcripts cargo to drive mesenchymal stem/stromal cells vasculogenic mimicry

doi: 10.1186/s12964-025-02401-x

Figure Lengend Snippet: VT107 alters the capacity of extracellular vesicles isolated from TNFα-treated U87 glioblastoma cells to trigger in vitro mesenchymal stem cells vasculogenic mimicry. ( A ) Representative phase contrast images were taken to monitor the formation of mesenchymal stem/stromal cells capillary-like structures for up to 6 h (upper panels) and were analyzed via Wimasis (lower panels). ( B ) Quantification from Wimasis analysis of the total loops over time. ( C ) Cultrex was depleted from soluble growth factors, then used to monitor the capacity to re-induce capillary-like structure formation upon 6 h (upper panels). The structures were analyzed via Wimasis (lower panels). Total loops were quantified to assess in vitro VM. ( D ) The impact of serum-free media (closed triangles), EVs (open circles) and of serum-enriched media (closed circles), or ( E ) the impact of EVs isolated from control, 30 ng/ml TNFα-, 1 µM VT107-, or TNFα/VT107-treated cells, was assessed on in vitro VM

Article Snippet: VM was assessed in vitro using Cultrex (3432-010-01, R&D Systems) to monitor capillary-like structures formation [ , ].

Techniques: Isolation, In Vitro, Control