mouse embryonic fibroblasts  (ATCC)

Journal: Foods

Article Title: Engineering a Compartmentalized Multi-Cell Co-Culture Hydrogel System Using Beeswax/Fucoidan/Alginate for Cultured Meat Modeling

doi: 10.3390/foods15101715

Figure Lengend Snippet: Cytotoxicity and cell attachment of C2C12 myoblasts through the Bw/Algi hydrogel. ( a ) Macroscopic appearance of the beeswax/alginate (Bw/Algi) hydrogel showing flexibility under compression. ( b , c ) Cytotoxicity assessment of Bw/Algi by CCK-8 assay and Live/Dead staining after indirect co-culture ( n = 4). The control group consisted of C2C12 cells cultured without Bw/Algi. Live cells are shown in green and dead cells in red. ( d ) SEM images showing C2C12 cell attachment on the surface of Bw/Algi hydrogels after 1 day of culture. White arrows indicate attached cells. Scale bar: 10 µm. ( e ) DAPI and Live/Dead staining showing C2C12 cells on Bw/Algi hydrogels after 3 days of culture. Scale bar: 200 µm. 3D Bw/Fu/Algi hydrogel discs were analyzed after proliferation/differentiation. Sections of Bw/Algi were obtained from both the central region (separating HECs and MEFs) and the peripheral region (separating MEFs and C2C12 cells). ( f ) After 3 days of proliferation, limited cell presence was observed at the central barrier, while no visible cells were observed at the peripheral barrier. The dashed box indicates the area shown at higher magnification. ( g ) After 7 days of differentiation, no visible cells were detected at either the central or peripheral Bw/Algi barrier. Scale bars = 200 and 100 μm. Data are presented as mean ± SE. Statistical analysis was performed using two-way ANOVA followed by Holm–Šidák multiple comparisons test; no significant differences were detected.

Article Snippet: Mouse C2C12 myoblasts (CRL-1772, ATCC, Manassas, VA, USA), mouse embryonic fibroblasts (MEF-1; CRL-2214, ATCC, Manassas, VA, USA), and EA.hy926 human endothelial cells (HECs) (ATCC, Manassas, VA, USA) were co-cultured under both 2D and 3D conditions.

Techniques: Cell Attachment Assay, CCK-8 Assay, Staining, Co-Culture Assay, Control, Cell Culture

Journal: Foods

Article Title: Engineering a Compartmentalized Multi-Cell Co-Culture Hydrogel System Using Beeswax/Fucoidan/Alginate for Cultured Meat Modeling

doi: 10.3390/foods15101715

Figure Lengend Snippet: Characterization of the 3D Bw/Fu/Algi hydrogel disc: differentiation, protein accumulation, and thermal behavior. ( a ) 3D co-culture disc showing C2C12 cells (1), MEFs (2), and HECs (3) embedded in Fu/Algi hydrogel and spatially separated by a Bw/Algi at day 3 of cell growth. ( b ) Immunofluorescence staining of skeletal myosin indicating myogenic marker expression in differentiated C2C12 cells after 7 days. A 2D co-culture conditions without hydrogel and mouse muscle tissue served as controls. Scale bar = 100 μm. ( c ) Total protein content measured using a BCA assay after 10 days of differentiation. ( d ) DSC thermograms showing thermal transitions of the hydrogel components compared with native bovine muscle. Data are presented as mean ± SE ( n = 3). Statistical significance was determined using one-way ANOVA: * p < 0.05, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Mouse C2C12 myoblasts (CRL-1772, ATCC, Manassas, VA, USA), mouse embryonic fibroblasts (MEF-1; CRL-2214, ATCC, Manassas, VA, USA), and EA.hy926 human endothelial cells (HECs) (ATCC, Manassas, VA, USA) were co-cultured under both 2D and 3D conditions.

Techniques: Co-Culture Assay, Immunofluorescence, Staining, Marker, Expressing, BIA-KA