Plant-Produced Human Recombinant Erythropoietic Growth Factors Support Erythroid Differentiation In Vitro

Abstract

Clinically available red blood cells (RBCs) for transfusions are at high demand, but in vitro generation of RBCs from hematopoietic stem cells requires significant quantities of growth factors. Here, we describe the production of four human growth factors: erythropoietin (EPO), stem cell factor (SCF), interleukin 3 (IL-3), and insulin-like growth factor-1 (IGF-1), either as non-fused proteins or as fusions with a carrier molecule (lichenase), in plants, using a Tobacco mosaic virus vector-based transient expression system. All growth factors were purified and their identity was confirmed by western blotting and peptide mapping. The potency of these plant-produced cytokines was assessed using TF1 cell (responsive to EPO, IL-3 and SCF) or MCF-7 cell (responsive to IGF-1) proliferation assays. The biological activity estimated here for the cytokines produced in plants was slightly lower or within the range cited in commercial sources and published literature. By comparing EC 50 values of plant-produced cytokines with standards, we have demonstrated that all four plant-produced growth factors stimulated the expansion of umbilical cord blood-derived CD34 + cells and their differentiation toward erythropoietic precursors with the same potency as commercially available growth factors. To the best of our knowledge, this is the first report on the generation of all key bioactive cytokines required for the erythroid development in a cost-effective manner using a plant-based expression system.

QR678 & QR678 Neo Hair Growth Formulations: A Cellular Toxicity & Animal Efficacy Study

“…It is a plant derivative consisting of biomimetic peptides, including Sh-Polypeptide 9 (biomimicking VEGF), Sh-Polypeptide 1 (biomimicking basic FGF), Sh-Oligopeptide 2 (biomimicking IGF-1), copper tripeptide-1, Sh-Polypeptide 3 (biomimicking KGF-1), Sh-Oligopeptide 4 (biomimicking thymosin β4), vitamins, minerals, and amino acids. 10 Although it has been proved that plant derivatives are biomimetic polypeptides of the growth factors and changing the source does not affect the efficacy of the products, 11 , 12 we wanted to confirm the same through our study. A similar animal and cytotoxicity trial was carried out using QR 678 Neo, and it was interesting to note that the results obtained were similar in both QR 678 and QR 678 Neo.…”

Estimation of the Changes in Hematological Standards of Albino Rabbits Treated with Different Concentrations for Alcoholic Extract of Cyperus rotundus (Nutsedge) and Rosmarinus officinalis (Rosemary)

“…For introduce an interpretation closer to the reality of the results obtained from the study, depending on the active compounds in the Nutsedge tubers and Rosemary leaves; Arginine, Glutamic and Aspartic acid produced an initial increase in hemoglobin level in the blood, so Threonine and Glutamic acid are an important amino acids that well contributes to the formation haemoglobin molecule and in the same time their dramatically diminishing and deprivation will lead to impairs of erythropoiesis and decreasing of Erythropoietin responsive cells in vitro (Musiychuk et al, 2013). Leucine and Isoleucine are abundant in hemoglobin 27% and would be expected to give a maximal response when added to the diet of rabbits daily for 8 weeks (Neuwirt and Ponka, 1977).…”

Plant‐derived human recombinant growth factors and serum albumin maintain stemness of human‐induced pluripotent stem cells

“…Recently, plant‐derived proteins have emerged as an alternative for biopharmaceuticals including vaccines, enzymes, antibodies, and therapeutic proteins because of their cost‐effective, high volume, low risk, and rapid expansion advantages (Buyel et al, 2017 ; Chen & Davis, 2016 ; Kunova et al, 2010 ; Moon et al, 2020 ; Musiychuk et al, 2013 ). Here, we show that plant‐derived growth factors allow to maintain the pluripotency of hiPSCs and to induce differentiation into NSCs.…”

“…Thus, the use of such growth factors may jeopardize the safety required for the commercialization of stem cell therapeutics. To solve these safety issues, a system for generating and using proteins produced in plants has been suggested as an alternative (Kunova et al, 2010 ; Musiychuk et al, 2013 ). Plant‐based proteins including vaccines, enzymes, antibodies, growth factors, and serum albumin have demonstrated the benefits such as being cost‐effective, having reduced risk, and allowing a rapid scale‐up (Buyel et al, 2017 ; Chen & Davis, 2016 ; Moon et al, 2020 ).…”