Journal: Pharmaceutics

Article Title: Melanoma Brain Metastases Patient-Derived Organoids: An In Vitro Platform for Drug Screening

doi: 10.3390/pharmaceutics16081042

Figure Lengend Snippet: Representative images of hematoxylin and eosin (H&E) staining and immunohistochemical staining (for Melan A, S100, HMB45, CD4, CD8, and Ki67) of the parental tumors and their MBM-PDO cultures (10× and 20× magnification).

Article Snippet: The MBM-PDO cultures were gently transferred from the ULA plates using a pipette, washed several times in basis medium, centrifuged, and resuspended in 3–5 mL of TrypLE Express (Gibco, ThermoFisher Scientific, Waltham, MA, USA) and incubated at 37 °C for 10 min. After washing the organoids with basis medium supplemented with 10% FCS, the cell pellet was resuspended in MBM-PDO culture medium, seeded in ultra-low attachment (ULA) plates (Stemcell Technologies, Vancouver, BC, Canada), and incubated at 37 °C.

Techniques: Staining, Immunohistochemical staining

Journal: Pharmaceutics

Article Title: Melanoma Brain Metastases Patient-Derived Organoids: An In Vitro Platform for Drug Screening

doi: 10.3390/pharmaceutics16081042

Figure Lengend Snippet: Bright-field images (5× magnification) and representative images of immunohistochemical stainings for Melan A and S100 staining (10× magnification) of different passages showing the stability of organoid-morphology and immunohistochemical profile. ( A ) SA34 MBM-PDO culture. ( B ) SA1 MBM-PDO culture. P, passage.

Article Snippet: The MBM-PDO cultures were gently transferred from the ULA plates using a pipette, washed several times in basis medium, centrifuged, and resuspended in 3–5 mL of TrypLE Express (Gibco, ThermoFisher Scientific, Waltham, MA, USA) and incubated at 37 °C for 10 min. After washing the organoids with basis medium supplemented with 10% FCS, the cell pellet was resuspended in MBM-PDO culture medium, seeded in ultra-low attachment (ULA) plates (Stemcell Technologies, Vancouver, BC, Canada), and incubated at 37 °C.

Techniques: Immunohistochemical staining, Staining

Journal: Pharmaceutics

Article Title: Melanoma Brain Metastases Patient-Derived Organoids: An In Vitro Platform for Drug Screening

doi: 10.3390/pharmaceutics16081042

Figure Lengend Snippet: ( A ) Graphical abstract for the in vitro drug sensitivity assay (created with Biorender.com). ( B ) A dose–response graph depicting the cell viability of MBM-PDO cultures treated with different concentrations of dabrafenib and trametinib for 72 h. Cell viability, assessed using the CellTiter-Glo assay (Promega), was normalized to the vehicle control. Each condition was tested in technical and biological triplicates. Data are presented as mean ± SEM. ( C ) Representative brightfield microscopy images of MBM-PDO cultures taken after 72 h of treatment with 1 µM and 0.125 µM dabrafenib and trametinib, as well as vehicle control (top row) (scale bar 100 µm).

Article Snippet: The MBM-PDO cultures were gently transferred from the ULA plates using a pipette, washed several times in basis medium, centrifuged, and resuspended in 3–5 mL of TrypLE Express (Gibco, ThermoFisher Scientific, Waltham, MA, USA) and incubated at 37 °C for 10 min. After washing the organoids with basis medium supplemented with 10% FCS, the cell pellet was resuspended in MBM-PDO culture medium, seeded in ultra-low attachment (ULA) plates (Stemcell Technologies, Vancouver, BC, Canada), and incubated at 37 °C.

Techniques: In Vitro, Sensitive Assay, Glo Assay, Control, Microscopy