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- Title
Ex Vivo Expansion of Functional Human UCB-HSCs/HPCs by Coculture with AFT024-hkirre Cells.
- Authors
Khan, Muti ur Rehman; Ali, Ijaz; Wei Jiao; Yun Wang; Masood, Saima; Yousaf, Muhammad Zubair; Javaid, Aqeel; Ahmad, Shafique; Meifu Feng
- Abstract
Kiaa1867 (human Kirre, hKirre) has a critical role in brain development and/or maintenance of the glomerular slit diaphragm in kidneys. Murine homolog of this gene, mKirre expressed in OP9 and AFT024 cells could support hematopoietic stem cells/hematopoietic progenitor cells (HSC/HPC) expansion in vitro. HKirre is also expressed in human FBMOB-hTERT cell line and fetal liver fibroblast-like cells but its function has remained unclear. In this paper,we cloned a hKirre gene fromhuman fetal liver fibroblast-like cells and established a stably overexpressing hKirre-AFT024 cell line. Resultant cells could promote self-renewal and ex vivo expansion of HSCs/HPCs significantly higher than AFT024-control cells transformed with mock plasmid. The Expanded human umbilical cord blood (hUCB) CD34+ cells retained the capacity of multipotent differentiation as long as 8 weeks and successfully repopulated the bone marrow of sublethally irradiated NOD/SCID mice, which demonstrated the expansion of longtermprimitive transplantable HSCs/HPCs. Importantly, hkirre could upregulate the expressions ofWnt-5A, BMP4, and SDF-1 and downregulate TGF-β with other hematopoietic growth factors. By SDS-PAGE andWestern Blot analysis, a ∼89 kDa protein in total lysate of AFT024-hKirre was identified. Supernatants from AFT024-hkirre could also support CD34+CD38- cells expansion. These results demonstrated that the AFT024-hKirre cells have the ability to efficiently expand HSCs/HPCs.
- Publication
BioMed Research International, 2014, Vol 2014, p1
- ISSN
2314-6133
- Publication type
Article
- DOI
10.1155/2014/412075