Date of Award
10-3-2019
Document Type
Thesis
Degree Name
Biology, MS
First Advisor
Jianfeng Xu
Committee Members
Anne Grippo; Maureen Dolan
Call Number
LD 251 . A566t 2019 W39
Abstract
Generation of red blood cells from hematopoietic stem cells (HSCs) used for blood transfusion represents one of the focus in regenerative medicine. Erythropoietic growth factors (eGFs), including stem cell factor (SCF), erythropoietin (EPO) and interleukin 3 (IL-3) are vital for proliferation and differentiation of HSCs into mature red blood cells (RBCs) ex vivo. However, massive production of RBCs requires significant quantity and high quality of eGFs, making manufacturing at large scale cost prohibitive. Plant cell culture is proposed to be a promising bioproduction platform for growth factors based on the advantages of, fast growing, no risk of human pathogen, and easy downstream separation and purification. However, low protein productivity is a common bottleneck towards commercialization of this production platform. This bottleneck is addressed by the proprietary “designer HypGP engineering” technology that leverages the plant-specific hydroxyproline (Hyp)-O-glycosylation code to design and engineer novel Hyp-O-glycosylated peptides (HypGP) in plant cells. The engineered HypGP can function as a molecular carrier to dramatically increase the secreted yields of fused proteins in plant cell culture. In this study, SCF were expressed in tobacco BY-2 cells with a HypGP tag engineered at both N-terminus and C-terminus. The secreted proteins were characterized and purified, and the yields were determined. The biological activities of the secreted tagged-SCF were assayed for their function in stimulating the proliferation of TF-1 cell line (human erythroblast cells), and also in facilitating the differentiation of HSC into RBC. Our research may provide a novel and promising plant cell-based platform to produce large quantity of eGFs that facilitate the hematopoietic stem cell research and clinical applications.
Rights Management
This work is licensed under a Creative Commons Attribution 4.0 International License.
Recommended Citation
Wang, Xiaoting, "Plant Cell-Derived Growth Factors for Ex Vivo Mass Production of Red Blood Cells" (2019). Student Theses and Dissertations. 395.
https://arch.astate.edu/all-etd/395
Included in
Biology Commons, Molecular, Cellular, and Tissue Engineering Commons, Plant Sciences Commons
