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Published on: September 22, 2013
[Test of transient expression of green fluorescent protein gene transferred to bone marrow stromal cells]
Xinquan Jiang1, Zhiyuan Zhang, Jun Cao
1Oral and Maxillofacial Department, Shanghai Ninth Hospital, Shanghai Second Medical University, Shanghai 200011, China.
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.
Conclusion:
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.
