[Influence of fluorescent protein expression on the proliferation of NIH3T3 cells in vitro].
OBJECTIVE
To investigate the influence of fluorescent protein expression on the proliferation of murine NIH3T3 cells, so as to provide a theoretical basis for cell tracing technology.


METHODS
NIH3T3 cells were cultured in vitro, and were randomly divided into control, pLEGFP-N1 (with transfection of pLEGFP-N1 retroviral vector), pEGFP-N1 (with transfection of pEGFP-N1 vector) and pDsRed2-C1 (with transfection of pDsRed2-C1 vector) groups. Then the cells were screened by G418 for 3 weeks. The changes in cell adhesive rate were observed and the population doubling times was determined by growth curve.


RESULTS
There was obvious fluorescent protein expression in the transfected NIH3T3 cells after G418 selection, and the highest percentage of labeled NIH3T3 cells was found in pLEGFP-N1 group. The population doubling time in pDsRed2-C1 (40.3+/-0.7 h) , PEGFP-N1 (39.6 +/- 0.6 h) and pLEGFP-N1 (36.5 +/- 0.7 h) groups was evidently longer than that in control (27.9 +/- 0.6 h, P < 0.01), with high adhesive rate in each group.


CONCLUSIONS
The expression of fluorescent protein exhibited some inhibitory effect on the proliferation of NIH3T3 cells in vitro. Since the inhibitory effect by retroviral vector was weaker compared with eukaryotic vector, it should be the first choice for fluorescent protein labeling during cell transplantation.
Duan
Xiao-jun
XJ
Center of Joint Surgery, Southwest Hospital, The Third Military Medical University, Chongqing 400038 , P. R. China.
Yang
Liu
L
Zhou
Yue
Y
Tang
Kang-lai
KL
Wang
Dong-wu
DW
D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D049452:Green Fluorescent Proteins / Q000096:biosynthesis*; D051379:Mice; D041681:NIH 3T3 Cells; D014162:Transfection
D049452:Green Fluorescent Proteins