Production of mouse granulocyte-macrophage colony-stimulating factor by gateway technology and transgenic rice cell culture

2011 ◽  
Vol 109 (5) ◽  
pp. 1239-1247 ◽  
Author(s):  
Yu-Kuo Liu ◽  
Li-Fen Huang ◽  
Shin-Lon Ho ◽  
Chun-Yu Liao ◽  
Hsin-Yi Liu ◽  
...  
Keyword(s):  
Cell Culture ◽  
Transgenic Rice ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Gateway Technology ◽  
Macrophage Colony ◽  
Stimulating Factor

scholarly journals Study on Hemostimulating Properties of Granulocyte-Macrophage Colony Stimulating Factor

2020 ◽  
Vol 20 (4) ◽  
pp. 268-276
Author(s):  
G. G. Shimina ◽  
A. V. Bateneva ◽  
S. G. Gamaley ◽  
T. I. Esina ◽  
T. G. Tereshchenko ◽  
...  
Keyword(s):  
Cell Culture ◽  
Proliferative Activity ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Dose Dependent ◽  
Stimulating Factor

The hemostimulating properties of granulocyte-macrophage colony-stimulating factor (GM-CSF) make possible its clinical use in alleviating side effects of anti-cancer radio- and chemotherapy, in bone marrow transplantation, and in the treatment of some primary immunodeficiency conditions associated with leukopenia. The State Research Center of Virology and Biotechnology “Vector” of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing has developed a high-performance technology for production of recombinant human GM-CSF (rhGM-CSF) based on a recombinant E. coli strain.The aim of the study was to assess hemostimulating activity of the rhGM-CSF preparation obtained using the new developed technology, as observed in cell culture and in the mice model of myelosuppression induced by cyclophosphamide administration.Materials and methods: in vitro evaluation of rhGM-CSF hemostimulating activity was performed by MTT assay in the commercial HL-60 promyelocytic leukemia cell culture with preliminary suppression of cell growth rate by adding a low concentration of dimethyl sulfoxide to the medium. In vivo studies were carried out in CBA/CaLac mice with cyclophosphamide-induced myelosuppression. The hemostimulating properties of the drug were evaluated after subcutaneous administration of 1–175 µg/kg doses for 4–5 days, following administration of a cytostatic agent. The total number of leukocytes and the content of their morphological forms were determined in blood samples taken at different time points after the drug administration. The statistical processing of the experimental data was based on analysis of variance using Statgraphics v. 5.0 software.Results: the proliferative activity of HL-60 cells incubated with the rhGM-CSF preparation for 72 hours was shown to be dose-dependent. The highest values of the increase in proliferative activity associated with an increase in the drug dose were observed in the concentration range from 0.04 to 0.64 ng/mL (proliferative activity increased by 11–18% when the dose was increased twofold). The experiments in mice demonstrated a two-phase pattern of the dose-dependent effect. The drug showed the highest hemostimulating effect at the dose of 90 µg/kg.Conclusions: the rhGM-CSF preparation obtained using the new developed technology has a pronounced hemostimulating activity confirmed by both in vitro and in vivo test systems. 

scholarly journals Enhancement of colony-forming activity of granulocyte-macrophage colony- stimulating factor by monocytes in vitro

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 918-924 ◽  
Author(s):  
M Namiki ◽  
H Hara
Keyword(s):  
Cell Culture ◽  
Single Cell ◽  
Colony Formation ◽  
Hematopoietic Progenitor Cell ◽  
Adherent Cells ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Single Cell Culture ◽  
Macrophage Colony ◽  
Stimulating Factor

Abstract Human recombinant granulocyte-macrophage colony-stimulating factor (hrGM-CSF) stimulated granulocyte-macrophage (GM) colony formation from human marrow mononuclear cells (MMCs) in a dose-dependent manner in methylcellulose culture. When phagocytes were depleted from MMCs, GM colony formation from the phagocyte-depleted (PD) MMCs by hrGM-CSF markedly decreased. Experiments in which PD-MMCs were cultured with hrGM-CSF and adherent cells showed that 94% (on day 7 and day 14) of the colonies from PD-MMCs were dependent on the presence of adherent cells. In contrast, the ability of granulocyte colony-stimulating factor (G-CSF) to form colonies was not affected by phagocyte depletion. To check for the presence or absence of progenitors that could form GM colonies in direct response to hrGM-CSF, single-cell culture of hematopoietic progenitor cell surface antigen (My-10)- positive PD-MMCs was carried out using a flow cytometer and an Autoclone System. In duplicate experiments, 0.7% and 3.5% (day 7) or 3.6% and 3.9% (day 14) of My-10-positive PD-MMCs formed GM colonies in response to hrGM-CSF and 5.1% and 6.0% (day 7) of My-10-positive PD- MMCs formed GM colonies in response to G-CSF. This was clear evidence for the presence of progenitors directly responding to hrGM-CSF. Also observed was a synergistic effect on GM colony formation in which more My-10-positive PD-MMCs stimulated by hrGM-CSF and G-CSF could form GM colonies than the sum of those stimulated by each separately. This enhancing effect of colony-forming activity of hrGM-CSF by adherent cells and the single cell culture experiment were reproduced in serum- free culture system.

scholarly journals Enhancement of colony-forming activity of granulocyte-macrophage colony- stimulating factor by monocytes in vitro

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 918-924
Author(s):  
M Namiki ◽  
H Hara
Keyword(s):  
Cell Culture ◽  
Single Cell ◽  
Colony Formation ◽  
Hematopoietic Progenitor Cell ◽  
Adherent Cells ◽  
Colony Stimulating Factor ◽  
Macrophage Colony Stimulating Factor ◽  
Single Cell Culture ◽  
Macrophage Colony ◽  
Stimulating Factor

Human recombinant granulocyte-macrophage colony-stimulating factor (hrGM-CSF) stimulated granulocyte-macrophage (GM) colony formation from human marrow mononuclear cells (MMCs) in a dose-dependent manner in methylcellulose culture. When phagocytes were depleted from MMCs, GM colony formation from the phagocyte-depleted (PD) MMCs by hrGM-CSF markedly decreased. Experiments in which PD-MMCs were cultured with hrGM-CSF and adherent cells showed that 94% (on day 7 and day 14) of the colonies from PD-MMCs were dependent on the presence of adherent cells. In contrast, the ability of granulocyte colony-stimulating factor (G-CSF) to form colonies was not affected by phagocyte depletion. To check for the presence or absence of progenitors that could form GM colonies in direct response to hrGM-CSF, single-cell culture of hematopoietic progenitor cell surface antigen (My-10)- positive PD-MMCs was carried out using a flow cytometer and an Autoclone System. In duplicate experiments, 0.7% and 3.5% (day 7) or 3.6% and 3.9% (day 14) of My-10-positive PD-MMCs formed GM colonies in response to hrGM-CSF and 5.1% and 6.0% (day 7) of My-10-positive PD- MMCs formed GM colonies in response to G-CSF. This was clear evidence for the presence of progenitors directly responding to hrGM-CSF. Also observed was a synergistic effect on GM colony formation in which more My-10-positive PD-MMCs stimulated by hrGM-CSF and G-CSF could form GM colonies than the sum of those stimulated by each separately. This enhancing effect of colony-forming activity of hrGM-CSF by adherent cells and the single cell culture experiment were reproduced in serum- free culture system.

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