Plant cell-secreted stem cell factor stimulates expansion and differentiation of hematopoietic stem cells. (January 2021)

• Record Type: Journal Article
• Authors: Wang, Xiaoting; Karki, Uddhab; Abeygunaratne, Hasara; UnnoldCofre, Carmela; Xu, Jianfeng
• Is Part Of: Process biochemistry. Volume 100(2021)
• Page Start: 39
• Page End: 48
• Publication Date: 2021-01
• Subjects: Plant cell culture -- Stem cell factor -- Hydroxyproline-O-glycosylation -- Hematopoietic stem cells -- Proliferation; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2020.09.029
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Plant cell culture is a promising production platform for high-quality growth factors. Hyp- O -glycosylated (SP)20 tag increases the secreted yields of SCF up to 2.5 μg/mL. The orientation of the (SP)20 tag substantially impacts the potency of SCF. (SP)20 -tagged SCF stimulates the differentiation of hematopoietic stem cell CD34 + cells. Abstract: Ex vivo generation of red blood cells (RBCs) from hematopoietic stem cells (HSCs) used for blood transfusion represents one of the focuses in current regenerative medicine. However, massive production of HSCs-based RBCs requires a significant quantity of erythropoietic growth factors, making manufacturing at large scale cost prohibitive. Plant cell culture is proposed to be a promising bioproduction platform for functional human proteins in a safe and cost-efficient manner. This study exploited a proprietary technology, named HypGP engineering technology, for high-yield production of one of the key erythropoietic growth factors--stem cell factor (SCF)--in plant cell culture. Specifically, a designer hydroxyproline (Hyp)- O -glycosylated peptide (HypGP) comprised of 20 tandem repeats of the "Ser-Pro" motif, or (SP)20, was engineered at either the N-terminus or C-terminus of SCF in tobacco BY-2 cells. The (SP)20 tag dramatically increased the secreted yields of SCF up to 2.5 μg/mL. The (SP)20 -tagged SCF showed bioactivity in promoting the proliferation of the TF-1 cell line, although the SCF-(SP)20 was 8.4-fold more potent than the (SP)20 -SCF. Both the (SP)20 -SCF and SCF-(SP)20 exhibited desired function in stimulating the expansion and differentiation of human umbilical cord blood CD34 + cells towards RBCs.