scholarly journals Purification of recombinant human fibroblast growth factor-2 from one-liter fermentation broth

2015 ◽  
Vol 18 (1) ◽  
pp. 14-22 ◽  
Author(s):  
Thy Tien Anh Tran ◽  
Trinh Thi My Nguyen ◽  
Hieu Van Tran
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Fermentation Broth ◽  
Cell Mass ◽  
Coli Strain ◽  
Fibroblast Growth Factor 2 ◽  
Fibroblast Growth ◽  
Multifunctional Protein ◽  
Dry Cell Weight ◽  
Dry Cell

Fibroblast Growth Factor - 2 (FGF-2), also known as basic FGF is a multifunctional protein that regulates the proliferation and differentiation of multiple types of cell. Recombinant human FGF-2 (rhFGF-2) is currently used in medicine, cosmetics, stem-cell culture, etc. In this study, we conducted one-liter scale fermentation using Escherichia coli strain that carries recombinant vector harboring FGF-2 coding gene to produce a large amount of FGF-2 protein. The evaluation of the fermentation efficacy was based on the growth curve, dry cell weight and amount of FGF-2 obtaining from one-liter fermentation. After fermentation, cell mass was lyzed by high pressure. Then, the FGF-2 in supernatant was purified by cation exchange and heparin-affinity chromatography. The purity and efficiency of the purification process were estimated by Bradford, silver staining and densitometry using QuantityOne software. The result showed that in oneliter fermentation, we obtained 5.2 g/liter dry cell-mass and 230 mg/liter FGF-2. The purity of FGF-2 was about 97.1 % and the purification efficiency was above 46.08 %.

scholarly journals Dynamic changes of podocytes caused by fibroblast growth factor 2 in culture

2021 ◽  
Author(s):  
Eishin Yaoita ◽  
Masaaki Nameta ◽  
Yutaka Yoshida ◽  
Hidehiko Fujinaka
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Fibroblast Growth Factor 2 ◽  
Quiescent State ◽  
Fibroblast Growth ◽  
Gene Expressions ◽  
Dynamic Changes ◽  
Ki 67

AbstractFibroblast growth factor 2 (FGF2) augments podocyte injury, which induces glomerulosclerosis, although the mechanisms remain obscure. In this study, we investigated the effects of FGF2 on cultured podocytes with interdigitating cell processes in rats. After 48 h incubation with FGF2 dynamic changes in the shape of primary processes and cell bodies of podocytes resulted in the loss of interdigitation, which was clearly shown by time-lapse photography. FGF2 reduced the gene expressions of constituents of the slit diaphragm, inflections of intercellular junctions positive for nephrin, and the width of the intercellular space. Immunostaining for the proliferation marker Ki-67 was rarely seen and weakly stained in the control without FGF2, whereas intensely stained cells were frequently found in the presence of FGF2. Binucleation and cell division were also observed, although no significant increase in cell number was shown. An in vitro scratch assay revealed that FGF2 enhanced migration of podocytes. These findings show that FGF2 makes podocytes to transition from the quiescent state into the cell cycle and change their morphology due to enhanced motility, and that the culture system in this study is useful for analyzing the pathological changes of podocytes in vivo.

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