Figure 1.
Figure 1.

Bovine stromal-vascular cells were grown from adipose tissue explants. Adipose tissue explants were compressed under coverslips held in place by a central drop of vacuum grease (A). Stromal-vascular cells were apparent at d 4 (B). Stromal-vascular cells continued to grow on the surface of the Petri dish and on the coverslip as shown on d 7 (C) and d 10 (D).

 


Figure 2.
Figure 2.

Differentiation of stromal-vascular cells into adipocytes. Stromal-vascular cells were induced to differentiate using differentiation media (Diff 1, 1.1, 2, 3) or grown in medium without differentiation components [Dulbecco’s modified Eagle’s medium (DMEM), Sigma-Aldrich, St. Louis, MO]. Differentiation measurements were executed on d 8 after initiation of differentiation and included incorporation of radiolabeled acetate into lipids (A) and activity of glycerol-3-phosphate dehydrogenase (G3PDH; B). The mRNA abundance of fatty acid-binding protein 4 (aP2; C), PPARγ (D), and acetyl-CoA carboxylase-α (ACCα; E) was measured by real-time PCR, with DMEM serving as the comparator. Error bars are SEM. Means lacking a common letter (a–c) are different (P < 0.05). This experiment was repeated 3 times with cells isolated from different animals, and duplicate wells were used for each measurement (n = 6).

 


Figure 3.
Figure 3.

Differentiation of stromal-vascular cells into adipocytes. Stromal-vascular cells were induced to differentiate using differentiation media (Diff 2, 2.1, 2.2, 2.3) or grown in medium without differentiation components [Dulbecco’s modified Eagle’s medium (DMEM), Sigma-Aldrich, St. Louis, MO]. Differentiation measurements were executed on d 8 after initiation and included incorporation of radiolabeled acetate into lipids (A) and activity of glycerol-3-phosphate dehydrogenase (G3PDH; B). The mRNA abundance of fatty acid-binding protein 4 (aP2; C), PPARγ (D), and acetyl-CoA carboxylase-α (ACCα; E) was measured by real-time PCR, with DMEM serving as the comparator. Error bars are SEM. Means lacking a common letter (a–d) are different (P < 0.05). This experiment was repeated 3 times with cells isolated from different animals, and duplicate wells were used for each measurement (n = 6).

 


Figure 4.
Figure 4.

Increasing seeding density increases adipocyte differentiation potential of stromal-vascular cells. Stromal-vascular cells were seeded at a low (6.7 × 103 cells/cm2) or high (2 × 104 cells/cm2) seeding density and cultured in the differentiation medium (Diff 2.2) or medium without differentiation components [Dulbecco’s modified Eagle’s medium (DMEM), Sigma-Aldrich, St. Louis, MO] for 8 d. Differentiation was assessed by incorporation of radiolabeled acetate into lipids. Error bars are SEM. Means lacking a common letter (a–c) are different (P < 0.05). This experiment was repeated 4 times with cells isolated from different animals, and duplicate wells were used for each measurement (n = 8).

 


Figure 5.
Figure 5.

Conjugated linoleic acid reduces adipocyte differentiation of bovine stromal-vascular cells. Stromal-vascular cells were induced to differentiate in the presence of BSA [no fatty acid (No FA)], 50 μM linoleic acid (18:2), or 50 μM trans-10, cis-12 CLA. Differentiation measurements were executed on d 8 after initiation and included incorporation of radiolabeled acetate into lipids (A). The mRNA abundance of fatty acid-binding protein 4 (aP2; B), PPARγ (C), acetyl-CoA carboxylase-α (ACCα; D), and stearoyl-CoA desaturase-1 (SCD1; E) was measured by real-time PCR, with No FA serving as the comparator. Error bars are SEM. Means lacking a common letter (a–c) are different (P < 0.05). This experiment was repeated 3 times with cells isolated from different animals (n = 3).

 


Figure 6.
Figure 6.

Conjugated linoleic acid reduces stearoyl-CoA desaturase-1 (SCD1) and acetyl-CoA carboxylase-α (ACCα) in bovine adipocytes differentiated from stromal-vascular cells. Stromal-vascular cells were induced to differentiate in the presence of BSA [no fatty acid (No FA)], 50 μM linoleic acid (18:2), or 50 μM trans-10, cis-12 CLA. The protein abundance of SCD1 and ACCα was determined by immunoblotting, with actin indicating equal loading. Error bars are SEM. Means lacking a common letter (a, b) are different (P < 0.05). This experiment was repeated 3 times with cells isolated from different animals, and pooled duplicate wells were used for each measurement (n = 3).