growth medium egm mv2  (PromoCell)

Journal: Advanced Science

Article Title: Subventricular Zone‐on‐a‐Chip: A Model to Study Neurogenesis Disruption in Neonatal Intraventricular Hemorrhage

doi: 10.1002/advs.202502145

Figure Lengend Snippet: Overview of the SVZ‐on‐a‐chip. A) The SVZ‐on‐a‐chip structure 24 h post‐seeding, showing distinct cellular compartments visualized via staining. The entire chip structure is shown (scale bar = 3000 µm) alongside a magnified maximum Z‐projection view (scale bar = 100 µm). Arrows indicate neural stem cells (NSCs), and triangles highlight human fetal astrocytes (HFAs) embedded in Matrigel, occupying the central channel. The right channel is lined with human brain microvascular endothelial cells (HBMECs), while the left channel contains human choroid plexus epithelial cells (HCPEpiC). B) Neural channel with HCPEpiC seeded along the central channel (scale bar = 200 µm). C) The central channel, highlighted by triangles, is shown in closer detail (scale bar = 200 µm). D) 3D view of the inner channel, showing cells stained with Nestin (gray). Astrocytes, marked by triangles, are stained red but lack Nestin expression (scale bar = 70 µm). E) Inner channel viewed from a different angle (scale bar = 70 µm). F) Vascular channel lined with HBMECs (scale bar = 70 µm). G) HBMECs fully covering the vascular channel (scale bar = 70 µm). H) A magnified maximum Z‐projection view of the SVZ‐on‐a‐chip 7 days after complete seeding, showing significant cellular growth (scale bar = 100 µm). Clusters of NSCs are observed invading both the neural and, predominantly, the vascular channel. I) Maximum Z‐projection view of Nestin staining in the middle channel (scale bar = 100 µm). J) Close‐up of DCX‐positive neural stem cells invading the vascular channel. DCX staining was used instead of Nestin because HBMECs also express Nestin (scale bar = 50 µm). SVZ: Subventricular Zone, NSCs: Neural Stem Cells, HFAs: Human Fetal Astrocytes, HBMECs: Human Brain Microvascular Endothelial Cells, HCPEpiC: Human Choroid Plexus Epithelial Cells, DCX: Doublecortin.

Article Snippet: DPBS minus Ca ++ and M g++ (14 190 144, Thermo Fischer, Waltham, Massachusetts, USA), DPBS with Ca ++ and Mg ++ (14 040 091, Thermo Fischer), Astrocyte Media (AM1801, ScienCell, Carlsbad, California, USA), TrypLE (12 604 013, Thermo Fischer), Attachment factor protein 1× (S006100, Thermo Fischer), EGM ‐2 MV Microvascular Endothelial Cell Growth Medium‐2 BulletKit (CC‐3202, Lonza, Basel, Switzerland), Endothelial Cell Growth Medium MV2 (C‐22022, PromoCell, Heidelberg, Germany), Epithelial Cell Medium (Innoprot, P60104 , Derio, Spain), poly‐L‐lysine (PLL) (Innoprot), DMEM: F12 Glutamax (31331‐028, Thermo Fischer), N2 supplement ( 17502‐048, Thermo Fischer), B27 serum free (11 530 536, Thermo Fischer), FGF (233‐FB, R&D Systems, MN, USA), EGF (E9644, Sigma Aldrich, Burlington, Massachusetts, USA), PLO (P3655, Sigma Aldrich), Laminin (L2020, Sigma Aldrich), Matrigel Growth Factor Reduced (GFR) Basement Membrane Matrix, LDEV‐free (354 230, Corning, Corning, New York, USA), Antibiotic‐Antimycotic (15 240 062, Themo Fischer), idenTx 3 Chip (Aim Biotech, Singapore), Transwells 0.4 μM microporous membrane (833 932 041, Sarstedt, Nümbrecht, Germany), High Pure RNA Isolation Kit (11 828 665 001, Roche, Basel, Switzerland), Clariom S Affymetrix Assay (902 927, Thermo Fischer), High‐capacity RNA‐to‐cDNA kit (4 387 406, Thermo Fischer), TaqMan probes (Applied Biosystems, California, USA), Fast Advanced Master Mix (4 444 557, Applied Biosystems), TRIzol (15 596 026, Thermo Fischer), Chloroform (194 002, MP Biomedicals, Santa Ana, California, USA), Cell recovery solution (354 253, Corning), Anti‐Adherence Rinsing Solution (0 7010, STEMCELL Technologies, Vancouver, Canada), AggreWell800 24‐well plates (34 811, STEMCELL Technologies), Goat serum (G9023, Sigma Aldrich), Trypan Blue (1 450 021, Bio‐Rad, Hercules, California, USA), LEGENDplex Human Inflammation Panel 1 (13‐plex) assay using a V‐bottom plate (BioLegend, San Diego, California, USA), CellROX Green reagent ( C10444 ,Invitrogen), JC‐1 dye (T3168, Invitrogen) IL1receptor antagonist (SRP3327, Sigma Aldrich).

Techniques: Staining, Expressing