J Korean Med Sci.  2009 Feb;24(1):104-109. 10.3346/jkms.2009.24.1.104.

Tissue Engineering of Injectable Soft tissue Filler: Using Adipose Stem Cells and Micronized Acellular Dermal Matrix

• Yoo G
• Lim JS

• KMID: 1794414
• DOI: http://doi.org/10.3346/jkms.2009.24.1.104

Abstract

In this study of a developed soft tissue filler, adipose tissue equivalents were constructed using adipose stem cells (ASCs) and micronized acellular dermal matrix (Alloderm). After labeling cultured human ASCs with fluorescent green protein and attaching them to micronized Alloderm (5X10(5) cells/1 mg), ASC-Alloderm complexes were cultured in adipogenic differentiation media for 14 days and then injected into the dorsal cranial region of nude male mice. The viabilities of ASCs in micronized Alloderm were determined at 1, 4, 7, and 14 days, and complexes, which had been cultured for 14 days and implanted in vivo for 2 months, were histologically evaluated by light, confocal, and scanning electron microscopy. The viabilities represented that ASCs in micronized Alloderm were alive during the culture period. ASC-Alloderm complexes cultured for 14 days contained round cells with large lipid vesicles by light microscopy and many spherical cells by SEM. ASCs in implanted ASCAlloderm complexes harvested from mice at 2 months postinjection were histologically found to have differentiated into adipocytes which had green fluorescence dye. Micronized Alloderm may be found useful as scaffold for human ASCs when constructing fat tissue for three-dimensional soft tissue filling. The present study suggests that ASC-Alloderm complexes can be used as injectable three-dimensional soft tissue fillers.

Keyword

Mesenchymal Stem Cells; Alloderm; Tissue Engineering

MeSH Terms

Adipocytes/*cytology
Adipogenesis
Adipose Tissue/cytology
Animals
Cell Differentiation
Cells, Cultured
Collagen/*chemistry
Fluorescent Dyes/chemistry
Injections, Subcutaneous
Male
Mice
Mice, Nude
Microscopy, Electron, Scanning
Stem Cell Transplantation/*methods
Stem Cells/cytology/pathology
Time Factors
Tissue Engineering/*methods
Transplantation, Heterologous

Figure

• 

  Fig. 1 Microscopic findings of adipose stem cells (ASCs) cultured in a monolayer for 14 days. (A) Some ASCs had differentiated into adipocytes, which contained many lipid droplets as visualized by Oil-red O staining. (B) ASCs were well labeled with PHK67 green fluorescent dye (×400).

• 

  Fig. 2 Changes in ASCs viabilities over 14 days in culture. ASCs were cultured in vitro after mixing then with micronized Alloderm (5×104 cells/1 mg of Alloderm). Cell viabilities were determined using XTT colorimetric assays. Values represent mean±standard deviation (error bar).

• 

  Fig. 3 Microscopic findings of in vitro cultured ASC-Alloderm complex for 14 days. ASCs in micronized Alloderm differentiated to round cells (arrows) with large lipid vesicles (H&E stain, ×400).

• 

  Fig. 4 Scanning microscopic findings after 14 day of in vitro culture. (A) Micronized Alloderm alone. (B) ASC-Alloderm complex. Many cells were found to cover the micronized Alloderm, and some spherical adipocyte-like cells were observed.

• 

  Fig. 5 Microscopic findings after 2 months of in vivo implantation. (A) Micronized Alloderm alone. Some fibroblast-like cells and few small capillaries were observed in the micronized Alloderm. (B) ASC-Alloderm complex. Many signet-ring cells and large capillaries were found (H&E stain, ×400).

• 

  Fig. 6 Confocal microscopic findings of injected ASC-Alloderm complexes excised at 2 months after implantation. Many signet-ring cells were stained by Oil-red O and some cells showed weak green fluorescence (Oil-red O stain, ×400).

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