De novo adipogenesis in mice at the site of injection of basement membrane and basic fibroblast growth factor - PubMed (original) (raw)

De novo adipogenesis in mice at the site of injection of basement membrane and basic fibroblast growth factor

N Kawaguchi et al. Proc Natl Acad Sci U S A. 1998.

Abstract

Autografting of fat pads has a long history in plastic and reconstructive surgery for augmentation of lost soft tissue. However, the results are disappointing because of absorption of the grafts with time. The fate of transplanted fat is linked to adipose precursor cells distributed widely in connective tissues. Adipocyte precursor cells can proliferate and mature into adipocytes even in the adult body depending on microenvironment. When reconstituted basement membrane, Matrigel, supplemented with more than 1 ng/ml bFGF was injected s.c. into 6-week-old mice, the neovascularization induced within 1 week was followed by migration of endogenous adipose precursor cells, and a clearly visible fat pad was formed. The pad grew until 3 weeks after the injection and persisted for at least 10 weeks. Such de novo adipogenesis was induced reproducibly by s.c. injection of Matrigel and bFGF over the chest, lateral abdomen, or head. Adipogenesis could be induced even in ear cartilage or in muscle. Thus, our results demonstrated that an abundant population of adipose precursor cells is distributed widely in connective tissues of the adult body and that they migrate into the neovascularized plug of Matrigel for proliferation and maturation. These results suggest a technique of augmenting lost soft tissue in plastic and reconstructive surgery.

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Figures

Figure 1

Figure 1

Enhanced adipogenesis of 3T3-F442A cells and de novo adipogenesis by Matrigel in combination with bFGF. 3T3-F442A preadipocytes were suspended in Matrigel with (D) or without (B) 1 μg/ml bFGF, and 100 μl of the cell suspension containing 2 × 106 cells was s.c. injected into a BALB/c nude mouse over the chest. As the control, 100 μl of Matrigel with (C) or without (A) 1 μg/ml bFGF also was injected. Arrows indicate fat pads formed 5 weeks after the injection.

Figure 2

Figure 2

Process of de novo adipogenesis by injection of Matrigel and bFGF. Matrigel was s.c. injected together with 1 μg/ml bFGF over the chests of 6-week-old mice, and well defined plugs between skin and muscle were excised for histological staining with hematoxylin and eosin. Neovascularization in Matrigel is completed within 1 week, and blood vessels with a clear endothelium lining are formed _(_A). The neovascularization was accompanied by invasion of fibroblast-like cells that differentiate into adipocytes within 2 weeks (B). The population and size of the adipocytes increased until 5 weeks after injection (C). The fat plugs formed de novo by injection of Matrigel and bFGF were preserved for 10 weeks (D) whereas the plugs formed by injecting Matrigel alone lacked many cells (E). (Bar = 100 μm.)

Figure 3

Figure 3

Lipid staining of plugs formed by Matrigel injection together with or without bFGF. Frozen sections of the plugs 5 weeks after injecting Matrigel together with (B) or without (A) bFGF were prepared and stained with Sudan IV followed by counterstaining in Harris hematoxylin. Other details as in Fig. 1. (Bar = 100 μm.)

Figure 4

Figure 4

Dependency of de novo adipogenesis on bFGF dose. Six-week-old mice were s.c. injected with Matrigel containing 0, 1, 10, 100, or 1,000 ng/ml bFGF (two injections into three mice) over the chest. (A) Averaged weight with SE of the Matrigel plugs excised 5 weeks after the injection. (B) Histological staining with hematoxylin and eosin of the tracts formed by injecting Matrigel containing 1 (a), 10 (b), 100 (c), or 1,000 (d) ng/ml bFGF. (Bar = 100 μm.)

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