PMID- 9603173
OWN - NLM
STAT- MEDLINE
DCOM- 19980619
LR  - 20190713
IS  - 0041-1337 (Print)
IS  - 0041-1337 (Linking)
VI  - 65
IP  - 9
DP  - 1998 May 15
TI  - Long-term expression of the gene encoding green fluorescent protein in murine 
      hematopoietic cells using retroviral gene transfer.
PG  - 1233-40
AB  - BACKGROUND: A major goal in retroviral-based gene therapy is to establish methods 
      that allow for selection and tracking of transduced cell populations. Green 
      fluorescent protein (GFP) may be useful for gene therapy applications because it 
      is a naturally fluorescent protein that can be detected using conventional flow 
      cytometers facilitating rapid analysis and purification of transduced cell 
      populations. However, it is unknown whether GFP can be stably expressed in vivo, 
      particularly in multiple bone marrow-derived cell lineages. METHODS: A murine 
      retrovirus carrying the gene encoding GFP was used to infect murine bone marrow 
      cells (BMCs). These studies were conducted to (1) directly determine whether GFP 
      could be used as a marker of BMC transduction, (2) determine whether GFP is 
      capable of being expressed in multiple bone marrow-derived hematopoietic cell 
      lineages, and (3) determine whether GFP could be used to follow the fate of 
      transduced cells in vivo. RESULTS: Infection of BMCs with retroviruses carrying 
      the gene encoding GFP resulted in a fluorescent signal in viable transduced cells 
      that was detectable by flow cytometry. Expression of GFP was detected in multiple 
      bone marrow-derived cell lineages after transduction, including stem cell 
      antigen-positive (Sca-1+), lineage marker-negative (Lin-) cells. Using GFP as a 
      selectable marker, we were able to enrich for transduced cells by cell sorting. 
      Mice reconstituted with enriched populations of GFP+ cells showed a significant 
      increase in the percentage of cells expressing GFP in the periphery when compared 
      with mice reconstituted with unenriched transduced bone marrow. CONCLUSIONS: 
      These data indicate that GFP can be used to select for transduced BMCs in vitro, 
      expressed in multiple bone marrow-derived cell lineages, used to select 
      transduced cells, and follow the fate of transduced cells long-term in vivo.
FAU - Bagley, J
AU  - Bagley J
AD  - Transplantation Biology Research Center, Molecular Neurogenetics Unit, 
      Massachusetts General Hospital and Harvard Medical School, Boston 02129, USA.
FAU - Aboody-Guterman, K
AU  - Aboody-Guterman K
FAU - Breakefield, X
AU  - Breakefield X
FAU - Iacomini, J
AU  - Iacomini J
LA  - eng
GR  - NS24279/NS/NINDS NIH HHS/United States
GR  - R01 HL48049-05/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Transplantation
JT  - Transplantation
JID - 0132144
RN  - 0 (Indicators and Reagents)
RN  - 0 (Luminescent Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/metabolism/virology
MH  - Cell Line/virology
MH  - Gene Expression/*physiology
MH  - *Gene Transfer Techniques
MH  - Green Fluorescent Proteins
MH  - Hematopoietic Stem Cells/*physiology/virology
MH  - *Indicators and Reagents
MH  - Luminescent Proteins/*genetics
MH  - Mice
MH  - Retroviridae/*genetics
MH  - Time Factors
MH  - Transduction, Genetic/physiology
MH  - Virus Assembly/genetics
EDAT- 1998/05/29 00:00
MHDA- 1998/05/29 00:01
CRDT- 1998/05/29 00:00
PHST- 1998/05/29 00:00 [pubmed]
PHST- 1998/05/29 00:01 [medline]
PHST- 1998/05/29 00:00 [entrez]
AID - 10.1097/00007890-199805150-00015 [doi]
PST - ppublish
SO  - Transplantation. 1998 May 15;65(9):1233-40. doi: 
      10.1097/00007890-199805150-00015.