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Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells

Dynamic culture protocols have recently emerged as part of (bone) tissue engineering strategies due to their ability to represent a more physiological cell environment in vitro. Here, we described how a perfusion flow induced by a simple bioreactor system improves proliferation and osteogenic commit...

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Autores principales: Pasini, Alice, Lovecchio, Joseph, Ferretti, Giulia, Giordano, Emanuele
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6525824/
https://www.ncbi.nlm.nih.gov/pubmed/31191662
http://dx.doi.org/10.1155/2019/1304194
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author Pasini, Alice
Lovecchio, Joseph
Ferretti, Giulia
Giordano, Emanuele
author_facet Pasini, Alice
Lovecchio, Joseph
Ferretti, Giulia
Giordano, Emanuele
author_sort Pasini, Alice
collection PubMed
description Dynamic culture protocols have recently emerged as part of (bone) tissue engineering strategies due to their ability to represent a more physiological cell environment in vitro. Here, we described how a perfusion flow induced by a simple bioreactor system improves proliferation and osteogenic commitment of human bone marrow stromal cells. L88/5 cells were cultured in poly(methyl methacrylate) custom-milled communicating well plates, in the presence of an osteogenic cocktail containing 1α,25-dihydroxyvitamin D3, L-ascorbic acid 2-phosphate, and β-glycerophosphate. The dynamic cell culture was maintained under perfusion flow stimulation at 1 mL/min for up to 4 days and compared with a static control condition. A cell viability assay showed that the proliferation associated with the dynamic cell culture was 20% higher vs. the static condition. A significantly higher upregulation of the osteogenic markers runt-related transcription factor 2 (RUNX2), collagen type I (COL1A1), osteocalcin (BGLAP), alkaline phosphatase (ALPL), and osteopontin (SPP1) was detected when the perfusion flow stimulation was administered to the cells treated with the osteogenic cocktail. An in silico analysis showed that in the dynamic cell culture condition (i) the shear stress in the proximity of the cell layer approximates 10(−3) Pa, (ii) the nutrient and the waste product concentration is more homogeneously distributed than in the static counterpart, and (iii) perfusion flow was associated with higher nutrient consumption. In summary, increased cell proliferation and enhanced early phenotype commitment indicate that dynamic cell culture conditions, delivered via bioreactor systems, produce an enhanced in vitro environment for both basic and translational research in tissue engineering and regenerative medicine.
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spelling pubmed-65258242019-06-12 Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells Pasini, Alice Lovecchio, Joseph Ferretti, Giulia Giordano, Emanuele Stem Cells Int Research Article Dynamic culture protocols have recently emerged as part of (bone) tissue engineering strategies due to their ability to represent a more physiological cell environment in vitro. Here, we described how a perfusion flow induced by a simple bioreactor system improves proliferation and osteogenic commitment of human bone marrow stromal cells. L88/5 cells were cultured in poly(methyl methacrylate) custom-milled communicating well plates, in the presence of an osteogenic cocktail containing 1α,25-dihydroxyvitamin D3, L-ascorbic acid 2-phosphate, and β-glycerophosphate. The dynamic cell culture was maintained under perfusion flow stimulation at 1 mL/min for up to 4 days and compared with a static control condition. A cell viability assay showed that the proliferation associated with the dynamic cell culture was 20% higher vs. the static condition. A significantly higher upregulation of the osteogenic markers runt-related transcription factor 2 (RUNX2), collagen type I (COL1A1), osteocalcin (BGLAP), alkaline phosphatase (ALPL), and osteopontin (SPP1) was detected when the perfusion flow stimulation was administered to the cells treated with the osteogenic cocktail. An in silico analysis showed that in the dynamic cell culture condition (i) the shear stress in the proximity of the cell layer approximates 10(−3) Pa, (ii) the nutrient and the waste product concentration is more homogeneously distributed than in the static counterpart, and (iii) perfusion flow was associated with higher nutrient consumption. In summary, increased cell proliferation and enhanced early phenotype commitment indicate that dynamic cell culture conditions, delivered via bioreactor systems, produce an enhanced in vitro environment for both basic and translational research in tissue engineering and regenerative medicine. Hindawi 2019-05-02 /pmc/articles/PMC6525824/ /pubmed/31191662 http://dx.doi.org/10.1155/2019/1304194 Text en Copyright © 2019 Alice Pasini et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Pasini, Alice
Lovecchio, Joseph
Ferretti, Giulia
Giordano, Emanuele
Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells
title Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells
title_full Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells
title_fullStr Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells
title_full_unstemmed Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells
title_short Medium Perfusion Flow Improves Osteogenic Commitment of Human Stromal Cells
title_sort medium perfusion flow improves osteogenic commitment of human stromal cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6525824/
https://www.ncbi.nlm.nih.gov/pubmed/31191662
http://dx.doi.org/10.1155/2019/1304194
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