Cargando…
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...
Autores principales: | , , , |
---|---|
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 |
_version_ | 1783419772558376960 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6525824 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Hindawi |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT pasinialice mediumperfusionflowimprovesosteogeniccommitmentofhumanstromalcells AT lovecchiojoseph mediumperfusionflowimprovesosteogeniccommitmentofhumanstromalcells AT ferrettigiulia mediumperfusionflowimprovesosteogeniccommitmentofhumanstromalcells AT giordanoemanuele mediumperfusionflowimprovesosteogeniccommitmentofhumanstromalcells |