Commitment of C3H10T1/2 pluripotent stem cells to the adipocyte lineage

Abstract

The increase of adipose tissue mass associated with obesity is due in part to an increase in the number of adipocytes. This hyperplasia results from recruitment of pluripotent stem cells present in the vascular stroma of adipose tissue. A model cell culture system has been developed that recapitulates this process both ex vivo and in vivo. After treatment of pluripotent C3H10T1͞2 stem cells with bone morphogenic protein 4 (BMP4) during proliferation followed by differentiation inducers at growth arrest, the cells synchronously enter S phase and undergo mitotic clonal expansion, a hallmark of preadipocyte differentiation. Upon exiting the cell cycle, these cells express adipocyte markers and acquire adipocyte characteristics at high frequency. C3H10T1͞2 cells treated with BMP4 in cell culture and implanted s.c. into athymic mice develop into tissue indistinguishable from adipose tissue in normal fat depots. We interpret the findings as evidence that BMP4 is capable of triggering commitment of pluripotent C3H10T1͞2 stem cells to the adipocyte lineage.adipose tissue ͉ differentiation ͉ bone morphogenic protein 4 ͉ mitotic clonal expansion ͉ development

A Lox/CHOP‐10 crosstalk governs osteogenic and adipogenic cell fate by MSCs

“…Several studies have demonstrated that BMP4 has a well‐established role in triggering commitment of mesenchymal stem cells into the osteogenic and adipogenic lineage 8, 9, 10, 15. In addition, BMP4‐regulated osteogenic and adipogenic lineage commitment of MSCs is mutually exclusive 8.…”

“…Based on our previous studies that knockdown of Lox disrupts the BMP4‐induced adipocyte lineage commitment from MSCs10 and a theoretical inverse relationship exists between osteogenic and adipogenic lineage commitment and differentiation of MSCs,6 we hypothesized that knockdown of Lox would enhance BMP4‐induced osteogenesis of MSCs. To explore this, C3H10T1/2 cells, a well‐characterized mesenchymal stem cells20 and faithful model of MSCs to study adipocytic and osteogenic commitment and differentiation both in vitro and in vivo,8, 9, 21 was used. As expected, two key osteogenic transcription factors, Runx2 and Osterix, were up‐regulated by BMP4, which were further enhanced by Lox knockdown at the committed stage (day 0) (Figure 1A‐C).…”

“…However, it has also been demonstrated that BMPs have pro‐adipogenic effects 8. For example, BMP2/4 can induce adipocytic lineage commitment of MSCs through activation of p38 Smad/MAPK signalling 9, 10, 11. In addition, other side effects, such as increased cancer risk related to higher BMP doses, have also been reported 12.…”

G0/G1 switch gene 2 has a critical role in adipocyte differentiation

“…26 However, this postconfluence mitosis has not been observed in mouse C3H10T1/2 or human preadipocytes. 8,27 It has been suggested that preadipocytes isolated from fat have already undergone the prerequisite clonal expansion in vivo, and thus further cell cycle events would not be observed in such cells. 4 Nevertheless, it is clear that some of the checkpoint proteins for mitosis are involved in regulation of adipogenesis.…”

Artemisinin derivatives prevent obesity by inducing browning of WAT and enhancing BAT function

“…In vitro differentiation of C3H10T1/2 cells can be divided into two phases: a commitment phase induced by BMP4 or BMP7 and a differentiation phase induced by a defined cocktail. BMP7 during the commitment phase drives C3H10T1/2 cells to develop into brown adipocytes [8], whereas BMP4 promotes adipogenesis [9]. We treated C3H10T1/2 cells with ATM throughout the BMP4-induced commitment phase and the differentiation phase (group b), or in the commitment phase only (group c), or in the differentiation phase only (group d; Supplementary information, Figure S1D (Cyto C) were upregulated by ATM in a dose-dependent manner ( Figure 1C).…”