Differentiation of embryonic stem cells into adipocytes in vitro

Abstract

Embryonic stem cells, derived from the inner cell mass of murine blastocysts, can be maintained in a totipotent state in vitro. In appropriate conditions embryonic stem cells have been shown to differentiate in vitro into various derivatives of all three primary germ layers. We describe in this paper conditions to induce differentiation of embryonic stem cells reliably and at high efficiency into adipocytes. A prerequisite is to treat early developing embryonic stem cell-derived embryoid bodies with retinoic acid for a precise period of time. Retinoic acid could not be substituted by adipogenic hormones nor by potent activators of peroxisome proliferator-activated receptors. Treatment with retinoic acid resulted in the subsequent appearance of large clusters of mature adipocytes in embryoid body outgrowths. Lipogenic and lipolytic activities as well as high level expression of adipocyte specific genes could be detected in these cultures. Analysis of expression of potential adipogenic genes, such as peroxisome proliferator-activated receptors gamma and delta and CCAAT/enhancer binding protein beta, during differentiation of retinoic acid-treated embryoid bodies has been performed. The temporal pattern of expression of genes encoding these nuclear factors resembled that found during mouse embryogenesis. The differentiation of embryonic stem cells into adipocytes will provide an invaluable model for the characterisation of the role of genes expressed during the adipocyte development programme and for the identification of new adipogenic regulatory genes.

The roles of Wnt antagonists Dkk1 and sFRP4 during adipogenesis of human adipose tissue‐derived mesenchymal stem cells

“…As in vitro systems, various pre-adipose cell lines and primary cultures of adipose-derived stromal vascular cells have been successfully employed for adipogenic differentiation studies (Kanazawa et al 2005;Choi et al 2007). In addition to these models, embryonic stem cells have been demonstrated to differentiate into mature adipocytes in vitro (Dani et al 1997;Li et al 2007). In this study, we demonstrated that hAMSCs are capable of differentiating into adipocyte, chondrocyte, osteocyte and neuronal-like cells; also, we demonstrated in vitro regulation of adipogenesis, using a hAMSC model.…”

“…2007). In addition to these models, embryonic stem cells have been demonstrated to differentiate into mature adipocytes in vitro (Dani et al . 1997; Li et al .…”

The KLF2 Transcription Factor Does Not Affect the Formation of Preadipocytes but Inhibits Their Differentiation into Adipocytes

“…KLF2 -/-ES cells were created using a two-step targeting procedure as described previously (14). The established protocol for EB culture and differentiation into adipocytes is described previously (23). In summary, KLF2 -/and KLF2 +/+ ES cells were grown in DMEM supplemented with 15% fetal calf serum on mitomycin-treated fibroblast cell layers.…”

Using Pluripotent Stem Cells to Understand Normal and Leukemic Hematopoietic Development

“… 44 , 45 EBs derived from human or murine ESCs have been used to study cell fate decisions and the differentiation of various cell types during embryogenesis as a good model for hematopoietic development. 46 , 47 …”