Figure 5. Porcine adipocytes grown to produce macroscale cultured fat.

(A) Pig DFAT cells cultured under adipogenic conditions containing 2% or 20% fetal bovine serum (FBS) for 30 days. Fat cells are stained for lipid using BODIPY (green) and for cell nuclei using Hoescht 33342 (blue). Scale bars 100 μm. (B) Lipid and cell number quantification of 30-day pig adipocytes, based on BODIPY and Hoescht 33342 staining, respectively. AU stands for arbitrary units. Results tagged with ‘a’ represent a difference of p≤0.0001, while ‘b’ represents p≤0.01. n=4 (technical) for both groups. (C, Left) Cell-free 1.6% alginate gels on the left. (C, Right) Porcine adipocytes (differentiated in 2% FBS media) mixed with alginate (1.6% final concentration) to form bulk cultured fat. Scale bar represents 1 mm. (D) Lipid staining images (BODIPY) of 30-day in vitro porcine adipocytes, juxtaposed with native adipocytes from a 97-day old (97D) pig, as well as a chicken and a cow. Scale bars represent 100 µm.

Figure 5—figure supplement 1. Pig DFAT cells cultured under adipogenic conditions with 0% fetal bovine serum (FBS) containing media for 30 days.

Cells were grown similarly to adipocytes cultured in adipogenic media containing 2% FBS, except the media did not contain GlutaMAX or FBS. The induction and lipid accumulation phases lasted 6 and 24 days, respectively. Lipid staining is BODIPY (green) and nuclei staining is Hoescht 33342 (blue). Scale bar represents 100 μm.