Journal: Biology Open

Article Title: Three-dimensional model of glioblastoma by co-culturing tumor stem cells with human brain organoids

doi: 10.1242/bio.056416

Figure Lengend Snippet: Workflow of the co-culture method. Schematic outline of the different steps of the protocol. (A) Human induced pluripotent stem cells (hiPSCs) are seeded on microfibers in neural induction media to form embryoid body-like aggregates. (B) Aggregates are then embedded in Matrigel droplets, which favor inversion of the polarity and protrusion of the neuroepithelium. (C) Subsequent removal of the Matrigel and culture in maturation media on an orbital shaker result in expansion and maturation of the organoids. (D) In parallel, patient-derived GSCs are expanded in 2D adherent cultures and (E) infected with lentiviruses to express the fluorescent histone marker H2B-GFP. (F) H2B-GFP GSCs are then co-cultured with human brain organoids in a low adherence 96-well plate for 24 h, generating glioblastoma-organoid co-cultures (GOC) (G).

Article Snippet: On day 10, medium was changed to maturation medium (Basal 2 plus Supplement E, STEMCELL Technologies, 08571).

Techniques: Co-Culture Assay, Derivative Assay, Infection, Marker, Cell Culture

Journal: Biology Open

Article Title: Three-dimensional model of glioblastoma by co-culturing tumor stem cells with human brain organoids

doi: 10.1242/bio.056416

Figure Lengend Snippet: Generation of cerebral organoids from different induced pluripotent stem cell (iPSC) lines. (A) Timeline of the differentiation process. (B–D) Induced pluripotent stem cell lines IMR-90 (B), FSPS-13B (C) and BobC (D) have been differentiated to generate cerebral organoids. Representative images are shown for the different stages of the protocol from at least three different experiments: B′,C′,D′, neural induction on microfibers; B″,C″,D″, neuroepithelial budding upon Matrigel encapsulation; B‴,C‴,D‴, organoid expansion and maturation (black arrows in D‴ indicate the smooth edges of BobC-derived organoids, which are failing to develop). (E–H) Differentiation of IMR-90 cells with (E) or without (F) microfibres showing neural induction (E,F, black arrowheads show the bright appearance of the neuroepithelium) and correct formation and expansion of the neuroepithelial bulges (G,H, black arrowheads in G indicate budding epithelial structures). EB: embryoid body. Scale bars: B–D 100 µm; B′–B‴,C′–C‴,D′–D‴ 1000 µm; E–H 100 µm.

Article Snippet: On day 10, medium was changed to maturation medium (Basal 2 plus Supplement E, STEMCELL Technologies, 08571).

Techniques: Encapsulation, Derivative Assay

Journal: Biology Open

Article Title: Three-dimensional model of glioblastoma by co-culturing tumor stem cells with human brain organoids

doi: 10.1242/bio.056416

Figure Lengend Snippet: 5 . Differentiation state of GSCs in organoid media. (A,B) Immunostaining for TUJ and SOX2 in 2D culture of GSCs grown in either growth media (A) or organoid maturation media (B). Scale bars: 100 µm. (C) Quantification of the percentage of SOX2+ cells that co-express TUJ. n =4 t -test *** P <0,001.

Article Snippet: On day 10, medium was changed to maturation medium (Basal 2 plus Supplement E, STEMCELL Technologies, 08571).

Techniques: Immunostaining