[Test of transient expression of green fluorescent protein gene transferred to bone marrow stromal cells].
OBJECTIVE
To determine the optimized condition under which BMP expression vector will be constructed to transfect bone marrow stromal cells (MSCs), plasmid vector coding enhanced green fluorescence protein (EGFP) gene pEGFP was transferred into MSCs. The transfer efficiency and transient expression were subsequently tested.


METHODS
pEGFP plasmid was amplified and tested by an enzyme cutting technique in vitro. MSCs, which were initially obtained from the bone marrow of rabbits, were cultured in vitro and transferred with pEGFP by means of lipofectamine media methods. The ratio of plasmid and lipofectamine was varied according to the experiment design. Transfer efficiency and transient expression were evaluated by fluorescent microscopy.


RESULTS
Transfer efficiency was correlated with the ratio of plasmid and lipofectamine. The expression of EGFP began in 24 hours after transferring, reached maximum in 48-72 hours and decreased in 1 week, however there remained a weak expression for more than 3 weeks.


CONCLUSIONS
The efficiency of transferring pEGFP into MSCs could achieve to 30% with proper ratio of plasmid and lipofactamine. pEGFP was an ideal transient expression vector for MSCs gene transference.
Jiang
Xinquan
X
Oral and Maxillofacial Department, Shanghai Ninth Hospital, Shanghai Second Medical University, Shanghai 200011, China.
Zhang
Zhiyuan
Z
Cao
Jun
J
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D019485:Bone Morphogenetic Proteins / Q000235:genetics; D002478:Cells, Cultured; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008055:Lipids / Q000235:genetics; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D014162:Transfection
D019485:Bone Morphogenetic Proteins; D008055:Lipids; C086724:Lipofectamine; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins