期刊
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
卷 160, 期 5, 页码 1415-1427出版社
SPRINGER
DOI: 10.1007/s12010-009-8603-2
关键词
Notch signaling; rhDSL; Expression; Purification; HS/PCs
资金
- Science & Technology Commission of Shanghai Municipality [075407071, 06dj14001]
The notch signaling pathway plays an important role in inhibiting cell differentiation and enhancing the repopulation capability of hematopoietic stem/progenitor cells. In this study, we developed rhDSL, a novel soluble form of Notch ligand Delta-like-1, which contains the DSL domain and the N-terminal sequence of the ligand, and investigated its function in ex vivo expansion of human umbilical cord blood (UCB)-primitive hematopoietic cells. The coding sequence for rhDSL was cloned into a pQE30 vector, and the recombinant rhDSL, fused with a 6x His tag, was expressed in Escherichia coli as inclusion bodies after isopropyl beta-d-thiogalactoside induction. After renaturing by dilutions, the protein was purified through anion exchange followed by affinity chromatography. The purity of rhDSL protein was more than 99% with very low endotoxin. In combination with human c-kit ligand, the effect of rhDSL on ex vivo expansion of UCB CD34(+) cells was found to be optimal at 1.5 mu g/ml of rhDSL. The rhDSL protein might therefore be a potential supplement for the expansion of UCB-primitive hematopoietic cells.
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