Effect of endothelial cell-targeted soluble Notch ligand hD1R protein on expansion and engraftment of cord blood hematopoietic stem/progenitor cells / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 885-890, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-242033
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effects of endothelial cell- targeted soluble Notch ligand hD1R protein on expansion and engraftment of cord blood hematopoietic stem/progenitor cell (CB HSPCs).</p><p><b>METHODS</b>Recombinant hD1R protein was first induced and purified. Human cord blood CD34⁺ cells were co-cultured on human umbilical vein endothelial cells (HUVECs) supplemented with a cocktail containing 5 types of human cytokines including TPO, SCF, FL, IL-6, IL-3 (5GF) and soluble hD1R. The expansion of CD34⁺ cells was tested under different culture conditions including PBS group (PBS replaces HUVEC), hD1R group, sup group (HUVEC supernatant replaces HUVEC), fix group (fixed HUVEC replaces HUVEC), Day 0 group (Control). Cell cycle and apoptosis of cultured cells were also analyzed. Their progeny expanded in PBS or hD1R group were transplanted into sublethally irradiated NOD/SCID mice. The percentages of human CD45⁺ (hCD45⁺) cells in the marrow of recipient mice were determined by FACS 12 weeks later.</p><p><b>RESULTS</b>hD1R induced more expansion in the total number of CD34⁺ cells cocultured with HUVECs plus 5GF, which was 87.50-fold increase compared to the Day 0 group, and 7.98-fold increase than that of PBS group. FACS analysis also showed that the percentage of CD34⁺ cells was 77.0% in G0/G1 phase in the hD1R group, which indicated that hD1R enhanced HSPCs expansion and inhibited apoptosis. Moreover, hD1R significantly promoted human HSPC engraftment after BM transplantation in irradiated mice.</p><p><b>CONCLUSION</b>The Notch-mediated ex vivo expansion system has been established and hD1R promoted expansion and engraftment of human CB HSPCs, which provided the evidence for further clinical application.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Hematopoietic Stem Cells
/
Cells, Cultured
/
Coculture Techniques
/
Antigens, CD34
/
Endothelial Cells
/
Allergy and Immunology
/
Fetal Blood
/
Membrane Proteins
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Hematology
Year:
2014
Type:
Article
Similar
MEDLINE
...
LILACS
LIS