PMID- 23643088
OWN - NLM
STAT- MEDLINE
DCOM- 20130619
LR  - 20181202
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 29
IP  - 3
DP  - 2013 Mar
TI  - [Effect of GFP-containing lentivirus infection on the expression of octamer 
      transcription factor 4 in human umbilical cord mesenchymal stem cells].
PG  - 292-6
AB  - OBJECTIVE: To infect human umbilical cord mesenchymal stem cells (hUCMSCs) with 
      lentivirus carrying green fluorescence protein (GFP) and observe its effect on 
      octamer binding transcription factor 4 (Oct4) expression. METHODS: Mesenchymal 
      stem cells isolated from human umbilical cord by tissue adherence method were 
      cultured and detected for the the surface markers by flow cytometry. HUCMSCs were 
      infected with lentivirus carrying GFP at different multiplicity of infection 
      (MOI). GFP expression efficiency was observed using the fluorescence microscope 
      and flow cytometry to obtain optimal MOI value. The experiment included 
      non-transfection group and the GFP transfection group. The effect of GFP 
      lentivirus on cell proliferation was evaluated by MTT. Oct4 expression in hUCMSCs 
      cultured in vitro continuously for 2 and 8 weeks was measured by quantitative 
      reverse transcriptase PCR (qRT-PCR) and immunofluorescence staining. RESULTS: 
      HUCMSCs displayed fusiform shape as fibroblasts in vitro. Flow cytometry revealed 
      that the third-passage cells highly expressed CD29, CD105 and CD90, and lowly 
      expressed CD34 and CD45. At 96 h after infection with GFP lentivirus (MOI=20), 
      GFP positive rate of hUCMSCs was more than 75%, the highest level. MTT showed 
      nearly no change in cell proliferation in the transfected group in comparison 
      with the non-transfection group (P>0.05). The relative expression of Oct4 
      detected by qRT-PCR in cells cultured for 2 and 8 weeks were 0.9075+/-0.0124 and 
      0.8600+/-0.0135, respectively. Immunofluorescence staining showed that Oct4 
      expression was localized in the nucleus of cells. CONCLUSION: Lentivirus-mediated 
      GFP gene can be expressed in hUCMSCs and has no significant effect on the 
      expression of Oct4.
FAU - Tang, Li
AU  - Tang L
AD  - Department of Cardiology, Chongqing Medical University, Chongqing, China.
FAU - Chang, Jing
AU  - Chang J
LA  - chi
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (Integrin beta1)
RN  - 0 (Octamer Transcription Factor-3)
RN  - 0 (POU5F1 protein, human)
RN  - 0 (Receptors, Tumor Necrosis Factor, Type I)
RN  - 0 (Thy-1 Antigens)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Cell Proliferation
MH  - Female
MH  - Gene Expression
MH  - Gene Expression Regulation
MH  - Green Fluorescent Proteins/*genetics
MH  - Humans
MH  - Infant, Newborn
MH  - Integrin beta1/metabolism
MH  - Lentivirus/*genetics
MH  - Mesenchymal Stem Cells/*metabolism
MH  - Octamer Transcription Factor-3/*genetics
MH  - Pregnancy
MH  - Receptors, Tumor Necrosis Factor, Type I/metabolism
MH  - Thy-1 Antigens/metabolism
MH  - Transfection/*methods
MH  - Umbilical Cord/*cytology
EDAT- 2013/05/07 06:00
MHDA- 2013/06/20 06:00
CRDT- 2013/05/07 06:00
PHST- 2013/05/07 06:00 [entrez]
PHST- 2013/05/07 06:00 [pubmed]
PHST- 2013/06/20 06:00 [medline]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Mar;29(3):292-6.