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Published on: December 13, 2012
[Expression of a human FL eukaryotic expressing plasmid mediated by lipofectamine in HFCL cells]
1Department of Hematology & Oncology, First Clinical College, Norman Bethune University of Medical Sciences, Changchun 130021, China.
Aim:
Bone marrow stromal cell line-HFCL were transfected with the recombinant eukaryotic expressing vector-pIRESlneo/hFL by using liposome-mediated gene transfer method and get a stable expression.
Methods:
HFCL cells were transfected with the recombinant eukaryotic expressing vector-pIRESlneo/hFL by using liposome lipofectamine. Integration of hFL in the genome, transcription of hFL mRNA and expression of hFL protein in the transfected HFCL cells were assayed by Southern blot, Northern blot, Western blot and ELISA, the experiment of the human umbilical blood CD34+ cell multiplication.
Results:
hFL cDNA was integrated into HFCL genome successfully, hFL mRNA was transcripted, hFL protein was expressed with (60.3 +/- 0.1) ng. 10(6) cell(-1) x d(-1) and the experiment of the human umbilical blood CD34+ cell multiplication shows that hFL has obvious biological activity in the supernatant.
Conclusion:
The recombinant plasmid is proved to be stably expressed in HFCL cells and obvious biological activity of hFL was detectable in the supernatant of the transfected cells.
