[A study on transfection of green fluorescence protein gene into human adipose stromal cells in vitro].
OBJECTIVE
To make a comparison on the efficiency of two methods for transfecting Green Fluorescence Protein gene into human adipose tissue-derived stromal cells, and to study the biological properties and multipotential differentiation of gene-transfected cells.


METHODS
The human subcutaneous adipose tissue was obtained, digested with one volume of collagenase type I, and then cultured with BGJb medium. After subculture and expansion, the human adipose tissue-derived stromal cells infected with Ad-GFP or liposome were observed and analyzed with fluorescence microscopy and flow cytometry to assess transfection efficiency. The growth curve of transfected adipose tissue-derived stromal cells was protracted. The adipose tissue-derived storomal cells were induced to differentiate into osteoblasts, and non-transfected cells were set as control.


RESULTS
42.5% +/- 1.5 of the human adipose tissue-derived stomal cells infected with Ad-GFP were found to express GFP at a level higher than that of the control of liposome (11.40%). Infected adipose tissue-derived stromal cells were noted to form mineralized nodes by the use of Alizarin Red stain.


CONCLUSIONS
The human adipose tissue-derived stromal cells infected with Ad-GFP can express higher level of GFP, and can maintain the ability of proliferation and differentiation as the non-infected human adipose tissue-derived stromal cells do. The infected adipose tissue-derived stromal cells with Ad-GFP can track the change of adipose tissue-derived stromal cells in the study of multipotential differentiation and can serve as cellular vehicles for systemic gene delivery.
Lin
Yun-teng
YT
Department of Oral and Mauillofacial Surgery, West China Stomatology Hospital, Sichussan University, Chengdu 610041, China.
Jing
Wei
W
Chen
Xi-zhe
XZ
Qiac
Ju
J
Li
Zhi-yong
ZY
Yan
Zheng-bin
ZB
Wu
Ling
L
Tian
Wei-dong
WD
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D010006:Osteoblasts / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D049452:Green Fluorescent Proteins