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Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation

AIMS: Here we introduce a wide and complex study comparing effects of growth factors used alone and in combinations on human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation. Certain ways of cell behaviour can be triggered by specific peptides – growth factors, influencing c...

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Autores principales: Hefka Blahnova, Veronika, Dankova, Jana, Rampichova, Michala, Filova, Eva
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The British Editorial Society of Bone and Joint Surgery 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7437520/
https://www.ncbi.nlm.nih.gov/pubmed/32864112
http://dx.doi.org/10.1302/2046-3758.97.BJR-2019-0183.R2
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author Hefka Blahnova, Veronika
Dankova, Jana
Rampichova, Michala
Filova, Eva
author_facet Hefka Blahnova, Veronika
Dankova, Jana
Rampichova, Michala
Filova, Eva
author_sort Hefka Blahnova, Veronika
collection PubMed
description AIMS: Here we introduce a wide and complex study comparing effects of growth factors used alone and in combinations on human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation. Certain ways of cell behaviour can be triggered by specific peptides – growth factors, influencing cell fate through surface cellular receptors. METHODS: In our study transforming growth factor β (TGF-β), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1), and vascular endothelial growth factor (VEGF) were used in order to induce osteogenesis and proliferation of hMSCs from bone marrow. These cells are naturally able to differentiate into various mesodermal cell lines. Effect of each factor itself is pretty well known. We designed experimental groups where two and more growth factors were combined. We supposed cumulative effect would appear when more growth factors with the same effect were combined. The cellular metabolism was evaluated using MTS assay and double-stranded DNA (dsDNA) amount using PicoGreen assay. Alkaline phosphatase (ALP) activity, as early osteogenesis marker, was observed. Phase contrast microscopy was used for cell morphology evaluation. RESULTS: TGF-β and bFGF were shown to significantly enhance cell proliferation. VEGF and IGF-1 supported ALP activity. Light microscopy showed initial extracellular matrix mineralization after VEGF/IGF-1 supply. CONCLUSION: A combination of more than two growth factors did not support the cellular metabolism level and ALP activity even though the growth factor itself had a positive effect. This is probably caused by interplay of various messengers shared by more growth factor signalling cascades. Cite this article: Bone Joint Res 2020;9(7):412–420.
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spelling pubmed-74375202020-08-28 Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation Hefka Blahnova, Veronika Dankova, Jana Rampichova, Michala Filova, Eva Bone Joint Res Bone Biology AIMS: Here we introduce a wide and complex study comparing effects of growth factors used alone and in combinations on human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation. Certain ways of cell behaviour can be triggered by specific peptides – growth factors, influencing cell fate through surface cellular receptors. METHODS: In our study transforming growth factor β (TGF-β), basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), insulin-like growth factor 1 (IGF-1), and vascular endothelial growth factor (VEGF) were used in order to induce osteogenesis and proliferation of hMSCs from bone marrow. These cells are naturally able to differentiate into various mesodermal cell lines. Effect of each factor itself is pretty well known. We designed experimental groups where two and more growth factors were combined. We supposed cumulative effect would appear when more growth factors with the same effect were combined. The cellular metabolism was evaluated using MTS assay and double-stranded DNA (dsDNA) amount using PicoGreen assay. Alkaline phosphatase (ALP) activity, as early osteogenesis marker, was observed. Phase contrast microscopy was used for cell morphology evaluation. RESULTS: TGF-β and bFGF were shown to significantly enhance cell proliferation. VEGF and IGF-1 supported ALP activity. Light microscopy showed initial extracellular matrix mineralization after VEGF/IGF-1 supply. CONCLUSION: A combination of more than two growth factors did not support the cellular metabolism level and ALP activity even though the growth factor itself had a positive effect. This is probably caused by interplay of various messengers shared by more growth factor signalling cascades. Cite this article: Bone Joint Res 2020;9(7):412–420. The British Editorial Society of Bone and Joint Surgery 2020-08-19 /pmc/articles/PMC7437520/ /pubmed/32864112 http://dx.doi.org/10.1302/2046-3758.97.BJR-2019-0183.R2 Text en © 2020 Author(s) et al. https://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (CC BY-NC-ND 4.0) licence, which permits the copying and redistribution of the work only, and provided the original author and source are credited.
spellingShingle Bone Biology
Hefka Blahnova, Veronika
Dankova, Jana
Rampichova, Michala
Filova, Eva
Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
title Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
title_full Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
title_fullStr Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
title_full_unstemmed Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
title_short Combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
title_sort combinations of growth factors for human mesenchymal stem cell proliferation and osteogenic differentiation
topic Bone Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7437520/
https://www.ncbi.nlm.nih.gov/pubmed/32864112
http://dx.doi.org/10.1302/2046-3758.97.BJR-2019-0183.R2
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