scholarly journals Effect of Whole Tissue Culture and Basic Fibroblast Growth Factor on Maintenance of Tie2 Molecule Expression in Human Nucleus Pulposus Cells

2021 ◽  
Vol 22 (9) ◽  
pp. 4723
Author(s):  
Kosuke Sako ◽  
Daisuke Sakai ◽  
Yoshihiko Nakamura ◽  
Jordy Schol ◽  
Erika Matsushita ◽  
...  
Keyword(s):  
Cell Culture ◽  
Tissue Culture ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Nucleus Pulposus ◽  
Primary Cell Culture ◽  
Primary Cell ◽  
Fibroblast Growth ◽  
Nucleus Pulposus Cells ◽  
Akt Inhibitor

Previous work showed a link between Tie2+ nucleus pulposus progenitor cells (NPPC) and disc degeneration. However, NPPC remain difficult to maintain in culture. Here, we report whole tissue culture (WTC) combined with fibroblast growth factor 2 (FGF2) and chimeric FGF (cFGF) supplementation to support and enhance NPPC and Tie2 expression. We also examined the role of PI3K/Akt and MEK/ERK pathways in FGF2 and cFGF-induced Tie2 expression. Young herniating nucleus pulposus tissue was used. We compared WTC and standard primary cell culture, with or without 10 ng/mL FGF2. PI3K/Akt and MEK/ERK signaling pathways were examined through western blotting. Using WTC and primary cell culture, Tie2 positivity rates were 7.0 ± 2.6% and 1.9 ± 0.3% (p = 0.004), respectively. Addition of FGF2 in WTC increased Tie2 positivity rates to 14.2 ± 5.4% (p = 0.01). FGF2-stimulated expression of Tie2 was reduced 3-fold with the addition of the MEK inhibitor PD98059 (p = 0.01). However, the addition of 1 μM Akt inhibitor, 124015-1MGCN, only reduced small Tie2 expression (p = 0.42). cFGF similarly increased the Tie2 expression, but did not result in significant phosphorylation in both the MEK/ERK and PI3K/Akt pathways. WTC with FGF2 addition significantly increased Tie2 maintenance of human NPPC. Moreover, FGF2 supports Tie2 expression via MEK/ERK and PI3K/Akt signals. These findings offer promising tools and insights for the development of NPPC-based therapeutics.

scholarly journals Fibrin-Hyaluronic Acid Hydrogel (RegenoGel) with Fibroblast Growth Factor-18 for In Vitro 3D Culture of Human and Bovine Nucleus Pulposus Cells

2019 ◽  
Vol 20 (20) ◽  
pp. 5036 ◽  
Author(s):  
Sonja Häckel ◽  
Mona Zolfaghar ◽  
Jie Du ◽  
Sven Hoppe ◽  
Lorin M. Benneker ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hyaluronic Acid ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Nucleus Pulposus ◽  
3D Culture ◽  
Fibroblast Growth ◽  
Wild Type ◽  
Nucleus Pulposus Cells ◽  
Hyaluronic Acid Hydrogel

We investigated the effects of a fibrin-hyaluronic acid hydrogel (FBG–HA) and fibroblast growth factor 18 (FGF-18) for nucleus pulposus (NP) regeneration. Healthy bovine (n = 4) and human degenerated NP cells (n = 4) were cultured for 14 days in FBG-HA hydrogel with FGF-18 (∆51-mutant or wild-type) in the culture medium. Gene expression, DNA content, and glycosaminoglycan (GAG) synthesis were evaluated on day 7 and 14. Additionally, histology was performed. Human NP cells cultured in FBG-HA hydrogel showed an increase in collagen type II (COL2) and carbonic anhydrase XII (CA12) gene expression after 14 or 7 days of culture, respectively. GAG release into the conditioned medium increased over 14 days. Healthy bovine NP cells showed increased gene expression of ACAN from day 7 to day 14. Wild type FGF-18 up-regulated CA12 gene expression of human NP cells. Histology revealed an increase of proteoglycan deposition upon FGF-18 stimulation in bovine but not in human NP cells. The FBG-HA hydrogel had a positive modulatory effect on human degenerated NP cells. Under the tested conditions, no significant effect of FGF-18 was observed on cell proliferation or GAG synthesis in human NP cells.

Fibroblast Growth Factor-2 Maintains the Differentiation Potential of Nucleus Pulposus Cells In Vitro

Spine ◽  
2007 ◽  
Vol 32 (5) ◽  
pp. 495-502 ◽  
Author(s):  
Tsung-Ting Tsai ◽  
Asha Guttapalli ◽  
Erbil Oguz ◽  
Lih-Huei Chen ◽  
Alexander R. Vaccaro ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Nucleus Pulposus ◽  
Fibroblast Growth Factor 2 ◽  
Fibroblast Growth ◽  
Nucleus Pulposus Cells ◽  
Differentiation Potential

scholarly journals Half-life modeling of basic fibroblast growth factor released from growth factor-eluting polyelectrolyte multilayers

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ivan Ding ◽  
Amy M. Peterson
Keyword(s):  
Cell Culture ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Half Life ◽  
Cell Types ◽  
Polyelectrolyte Multilayers ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fibroblast Growth ◽  
Cell Culture Study

AbstractGrowth factor-eluting polymer systems have been widely reported to improve cell and tissue outcomes; however, measurements of actual growth factor concentration in cell culture conditions are limited. The problem is compounded by a lack of knowledge of growth factor half-lives, which impedes efforts to determine real-time growth factor concentrations. In this work, the half-life of basic fibroblast growth factor (FGF2) was determined using enzyme linked immunosorbent assay (ELISA). FGF2 release from polyelectrolyte multilayers (PEMs) was measured and the data was fit to a simple degradation model, allowing for the determination of FGF2 concentrations between 2 and 4 days of culture time. After the first hour, the FGF2 concentration for PEMs assembled at pH = 4 ranged from 2.67 ng/mL to 5.76 ng/mL, while for PEMs assembled at pH = 5, the concentration ranged from 0.62 ng/mL to 2.12 ng/mL. CRL-2352 fibroblasts were cultured on PEMs assembled at pH = 4 and pH = 5. After 2 days, the FGF2-eluting PEM conditions showed improved cell count and spreading. After 4 days, only the pH = 4 assembly condition had higher cells counts, while the PEM assembled at pH = 5 and PEM with no FGF2 showed increased spreading. Overall, the half-life model and cell culture study provide optimal concentration ranges for fibroblast proliferation and a framework for understanding how temporal FGF2 concentration may affect other cell types.

DNA aptamer assemblies as fibroblast growth factor mimics and their application in stem cell culture

2019 ◽  
Vol 55 (18) ◽  
pp. 2672-2675 ◽  
Author(s):  
Ryosuke Ueki ◽  
Saki Atsuta ◽  
Ayaka Ueki ◽  
Junya Hoshiyama ◽  
Jingyue Li ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Basic Fibroblast Growth Factor ◽  
Dna Aptamer ◽  
Fibroblast Growth ◽  
Single Stranded Dna ◽  
Stem Cell Culture

A 76-mer single-stranded DNA acts as functional mimic of basic fibroblast growth factor (bFGF), one of the essential factors for stem cell culture.

scholarly journals A Stable Chimeric Fibroblast Growth Factor (FGF) Can Successfully Replace Basic FGF in Human Pluripotent Stem Cell Culture

PLoS ONE ◽  
2015 ◽  
Vol 10 (4) ◽  
pp. e0118931 ◽  
Author(s):  
Yasuko Onuma ◽  
Kumiko Higuchi ◽  
Yasuhiko Aiki ◽  
Yujing Shu ◽  
Masahiro Asada ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Pluripotent Stem Cell ◽  
Fibroblast Growth ◽  
Human Pluripotent Stem Cell ◽  
Basic Fgf ◽  
Stem Cell Culture

scholarly journals αvβ3Integrin Mediates the Cell-adhesive Capacity and Biological Activity of Basic Fibroblast Growth Factor (FGF-2) in Cultured Endothelial Cells

1997 ◽  
Vol 8 (12) ◽  
pp. 2449-2461 ◽  
Author(s):  
Marco Rusnati ◽  
Elena Tanghetti ◽  
Patrizia Dell’Era ◽  
Anna Gualandris ◽  
Marco Presta
Keyword(s):  
Tissue Culture ◽  
Endothelial Cells ◽  
Cell Adhesion ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Amino Acid Sequences ◽  
Dependent Manner ◽  
Fibroblast Growth ◽  
Tissue Culture Plastic ◽  
Cell Adhesive

Fibroblast growth factor-2 (FGF-2) immobilized on non-tissue culture plastic promotes adhesion and spreading of bovine and human endothelial cells that are inhibited by anti-FGF-2 antibody. Heat-inactivated FGF-2 retains its cell-adhesive activity despite its incapacity to bind to tyrosine-kinase FGF receptors or to cell-surface heparan sulfate proteoglycans. Recombinant glutathione-S-transferase-FGF-2 chimeras and synthetic FGF-2 fragments identify two cell-adhesive domains in FGF-2 corresponding to amino acid sequences 38–61 and 82–101. Both regions are distinct from the FGF-receptor-binding domain of FGF-2 and contain a DGR sequence that is the inverse of the RGD cell-recognition sequence. Calcium deprivation, RGD-containing eptapeptides, soluble vitronectin (VN), but not fibronectin (FN), inhibit cell adhesion to FGF-2. Conversely, soluble FGF-2 prevents cell adhesion to VN but not FN, thus implicating VN receptor in the cell-adhesive activity of FGF-2. Accordingly, monoclonal and polyclonal anti-αvβ3antibodies prevent cell adhesion to FGF-2. Also, purified human αvβ3binds to immobilized FGF-2 in a cation-dependent manner, and this interaction is competed by soluble VN but not by soluble FN. Finally, anti-αvβ3monoclonal and polyclonal antibodies specifically inhibit mitogenesis and urokinase-type plasminogen activator (uPA) up-regulation induced by free FGF-2 in endothelial cells adherent to tissue culture plastic. These data demonstrate that FGF-2 interacts with αvβ3integrin and that this interaction mediates the capacity of the angiogenic growth factor to induce cell adhesion, mitogenesis, and uPA up-regulation in endothelial cells.

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