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Journal: iScience
Article Title: A simplified and defined serum-free medium for cultivating fat across species
doi: 10.1016/j.isci.2022.105822
Figure Lengend Snippet: rDM outperforms cDM in both 3%FBS and DMAD during long-term (28-day) 3D culture (A) Illustration of the experimental design. Bovine SVC were encapsulated with alginate hydrogel and differentiated with 3%FBS or DMAD. cDM contained all 4 inducers during first media change followed by progression for the whole period of culture, rDM contained rosiglitazone and insulin for 28 days. DMAD contained HC/PR. (B) Representative images of maximum intensity projection confocal microscopy. Differentiation with control (cDM) or reduced (rDM) differentiation medium in 3%FBS or DMAD at day 28. Blue, Hoechst; and yellow, Nile Red. (C) PLIN1 immunohistochemistry of 3D hydrogel/cell constructs differentiated in cDM or rDM medium at day 28. Blue, Hoechst; red, PLIN1; and yellow, Nile red. (B and C) Scale bar, 100 μm. (D) Quantitative analysis of glycerol release at day 0, 7 and 28. Data is representative of 3 independent experiments using 4 donors. (E) Qualitative and quantitative assessment of the amount of fatty acids within triglycerides (normalized to the amount of DNA). Results are representative of 5 independent experiments using 6 donors. (D and E) Statistical differences were analyzed with a one-way ANOVA. Data are represented as mean ± SD; NS, not significant; ∗∗∗p < 0.001. See also Figure S5 and .
Article Snippet: anti-mouse PLIN1 , Cell signalling technology , Rabbit mAb Cat #9349; RRID: AB_10829911.
Techniques: Confocal Microscopy, Control, Immunohistochemistry, Construct