Low temperature exposure induces browning of bone marrow stem cell derived adipocytes in vitro

Brown and beige adipocytes are characterised as expressing the unique mitochondrial uncoupling protein (UCP)1 for which the primary stimulus in vivo is cold exposure. The extent to which cold-induced UCP1 activation can also be achieved in vitro, and therefore perform a comparable cellular function, is unknown. We report an in vitro model to induce adipocyte browning using bone marrow (BM) derived mesenchymal stem cells (MSC), which relies on differentiation at 32°C instead of 37°C. The low temperature promoted browning in adipogenic cultures, with increased adipocyte differentiation and upregulation of adipogenic and thermogenic factors, especially UCP1. Cells exhibited enhanced uncoupled respiration and metabolic adaptation. Cold-exposed differentiated cells showed a marked translocation of leptin to adipocyte nuclei, suggesting a previously unknown role for leptin in the browning process. These results indicate that BM-MSC can be driven to forming beige-like adipocytes in vitro by exposure to a reduced temperature. This in vitro model will provide a powerful tool to elucidate the precise role of leptin and related hormones in hitherto functions in the browning process

MORPHOMETRIC-STEREOLOGIC ANALYSIS OF BROWN ADIPOCYTE DIFFERENTIATION IN ADULT MICE

Brown adipocyte differentiation from interstitial stem cells was analyzed by morphometric-stereologic methods in adult mice. Confirming previous studies, four different stages of development were identified: 1) interstitial cells → 2) protoadipocytes (interstitial cells with tiny lipid droplets) → 3) preadipocytes → 4) mature brown adipocytes. Brown adipocyte precursor cells (interstitial cells and protoadipocytes) occupied only a small fraction of total brown adipose tissue (BAT) volume (1.7 and 1.8%, respectively). Most of the BAT volume was occupied by fully differentiated multilocular cells (65% vol/vol), preadipocytes (12%), and blood capillaries (10%). The differentiation of protoadipocytes into preadipocytes was characterized by a doubling in the cellular volume (from 800 to 1,500 μm3) that was associated with a fivefold increase in the number of mitochondria (221 to 1,464), an eightfold augmentation in mitochondrial size (from 0.042 to 0.37 μm3), a fourfold increase in the surface density of the inner mitochondrial membrane (from 8 to 35 μm2/μm3), resulting in a 14-fold enlargement in the relative volume of the mitochondrial compartment (from 2 to 29%). This remarkable mitochondrial proliferation was accompanied by an increase in the number and volume of cytosolic lipid droplets. In contrast, the differentiation of preadipocytes into brown adipocytes was mainly characterized by a doubling in the size of the lipid compartment; the mean volume of single droplets increased 35 times but their number decreased 6-7 times. The mitochondrial modifications were minor; there was only a slight increase in the surface density of the inner membrane. In conclusion, the major step of brown adipocyte differentiation consists in the transformation of protoadipocytes into preadipocytes. It is characterized by a large proliferation of mitochondria with tightly packed cristae that is associated with a marked lipogenesis resulting in a significant expansion of the cellular volume