Ex vivo expansion of functional human UCB-HSCs/HPCs by coculture with AFT024-hkirre cells.
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 from human 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 long-term primitive transplantable HSCs/HPCs. Importantly, hkirre could upregulate the expressions of Wnt-5A, BMP4, and SDF-1 and downregulate TGF- β with other hematopoietic growth factors. By SDS-PAGE and Western 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.
Department of Pathology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
Muti ur Rehman
Mu
Khan
Institute of Biotechnology and Genetic Engineering, University of Agriculture, Peshawar 25000, Pakistan.
Ijaz
I
Ali
State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Wei
W
Jiao
State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Yun
Y
Wang
Department of Anatomy and Histology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
Saima
S
Masood
Department of Advanced Materials & Nanotechnology, College of Engineering, Peking University, Beijing 100871, China.
Muhammad Zubair
MZ
Yousaf
Department of Pharmacology and Toxicology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
Aqeel
A
Javaid
Institue of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.
Shafique
S
Ahmad
State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
Meifu
M
Feng
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D049109:Cell Proliferation; D018920:Coculture Techniques; D005312:Fetal Blood / Q000166:cytology; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice
C474214:Kirrel3 protein, mouse; D008565:Membrane Proteins