Adipose tissues differentiated by adipose-derived stem cells harvested from transgenic mice.
OBJECTIVE
To induce adipocyte differentiation in vitro by adipose-derived stromal cells (ASCs) harvested from transgenic mice with green fluorescent protein (GFP)and assess the possibility of constructing adipose tissues via attachment of ASCs to type I collagen scaffolds.


METHODS
Inguinal fat pads from GFP transgenic mice were digested by enzymes for isolation of ASCs (primary culture). After expansion to three passages of ASCs, the cells were incubated in an adipogenic medium for two weeks, and the adipocyte differentiation by ASCs in vitro was assessed by morphological observation and Oil Red O staining. Then they were attached to collagen scaffolds and co-cultured for 12 hours, followed by hypodermic implantation to the dorsal skin of nude mice for 2 months. The newly-formed tissues were detected by HE staining.


RESULTS
The cultured primary stem cells were fibroblast-like and showed active proliferation. After being incubated in an adipocyte differentiation medium, the lipid droplets in the cytoplasm accumulated gradually and finally developed into mature adipocytes, which showed positive in Oil Red O staining. A 0.5-cm3 new tissue clot was found under the dorsal skin of the nude mice and it was confirmed as mature adipose tissues by fluorescent observation and HE staining.


CONCLUSIONS
ASCs can successfully differentiate adipose tissues into mature adipocytes, which exhibit an adipocyte-like morphology and express as intracytoplasmic lipid droplets. It is an efficient model of adipose tissues engineered with ASCs and type I collagen scaffolds.
Lu
Feng
F
Department of Plastic and Reconstructive Surgery, Nanfang Medical University, Guangzhou 510515, China. lufengmail2005@hotmail.com
Gao
Jian-hua
JH
Ogawa
Rei
R
Mizuro
Hiroshi
H
Hykusoku
Hiki
H
D050156:Adipogenesis; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D013234:Stem Cells / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology; D023822:Tissue Engineering / Q000379:methods*