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: PLoS ONE

Article Title: Cell Population Kinetics of Collagen Scaffolds in Ex Vivo Oral Wound Repair

doi: 10.1371/journal.pone.0112680

Figure Lengend Snippet: The ex vivo wound healing model is shown in top view and cross-sectional view (A). The bottom of the well was covered with an acellular growth factor reduced basal membrane extract (BME). The scaffold (sponge type scaffold composed of cross-linked collagen or the gel type scaffold composed of rat collagen I) in the center was surrounded by a BME-gel containing 800,000 hGFs/ml and covered with an acellular BME gel and Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 1% fetal bovine serum (FBS) and antibiotics. Both scaffold and gel were tested with and without the addition of platelet-derived growth factor-BB (PDGF). Fluorescence images were taken to assess DiI-labeled cells within the scaffold over time. To assess the metabolic activity and gene expression dynamics, the scaffolds including the “active zone” (dashed line) were subjected to MTT tests, gene array analysis, and quantitative PCR. (B) Representative scanning electron microscopy images depict the morphology of the collagen gel, collagen scaffold at 400×, and 800× magnification. The white bar represents 100 µm.

Article Snippet: 48 well plates were first layered with 50 μL of 12 mg/ml growth factor reduced basal membrane extract (BME; CULTREX, TREVIGEN, Gaithersburg, MD, USA) without cells.

Techniques: Ex Vivo, Membrane, Modification, Derivative Assay, Fluorescence, Labeling, Activity Assay, Gene Expression, Real-time Polymerase Chain Reaction, Electron Microscopy