[Adipocytes differentiation of NIH3T3 cells induced by peroxisome proliferator activation receptor gamma 2 expression].
OBJECTIVE
To express the mouse peroxisome proliferator activated receptor gamma 2(mPPAR gamma 2) in NIH3T3 cells mediated by the recombinant retrovirus and study its function.


METHODS
mPPAR gamma 2 gene digested from the recombinant plasmid pcDNA3/mPPAR gamma 2 and confirmed to contain the target gene segment with fluorescence-sequencing was subcloned into retrovirus vector pGCEN to generate the recombinant retrovirus pGCEN/mPPAR gamma 2. The recombinant retrovirus pGCEN/mPPAR gamma 2 and pGCEN were packaged with PA317 cells and anti-G418 clones of PA317 cells were selected. Viral supernatants were collected and used to infect NIH3T3 cells. Peroxisome proliferator activated receptor gamma 2 (PPAR gamma 2)-expressing NIH3T3 cells cultured in the differentiation media containing PPAR gamma activator ETYA were induced into adipocytes.


RESULTS
The recombinant retrovirus pGCEN/mPPAR gamma 2 was constructed, 5 x 10(4) CFU/ml of the viral supernatants containing pGCEN/mPPAR gamma 2 and 6 x 10(5) CFU/ml of the viral supernatants containing pGCEN were obtained. mPPAR gamma 2 was expressed in NIH3T3 cells mediated by the recombinant retrovirus. Lipid accumulation obviously existed in PPAR gamma 2-expressing NIH3T3 cells at 10 days postdifferentiation and the lipid-containing cells morphologically resembled the mature adipocytes in vivo.


CONCLUSIONS
An adipocyte differentiation model in vitro was established. The work is the basis for further researches on the molecular mechanism of adipocyte differentiation induced by PPAR gamma 2.
Zuo
X
X
Shanghai Ruijin Hospital, Shanghai Institute of Endocrinology, Shanghai the Second Medical University, Shanghai 200025, China.
Li
G
G
Luo
T
T
Li
J
J
Liu
Y
Y
Sun
W
W
Luo
M
M
D016475:3T3 Cells / Q000166:cytology / Q000378:metabolism*; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D066298:In Vitro Techniques; D051379:Mice; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis* / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics
D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors