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Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures
INTRODUCTION: Recent studies in the literature have highlighted the critical role played by cell signalling in determining haemopoietic stem cell (HSC) fate within ex vivo culture systems. Stimulatory signals can enhance proliferation and promote differentiation, whilst inhibitory signals can signif...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4443622/ https://www.ncbi.nlm.nih.gov/pubmed/25888759 http://dx.doi.org/10.1186/s13287-015-0048-7 |
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author | Berry, Joseph D Godara, Pankaj Liovic, Petar Haylock, David N |
author_facet | Berry, Joseph D Godara, Pankaj Liovic, Petar Haylock, David N |
author_sort | Berry, Joseph D |
collection | PubMed |
description | INTRODUCTION: Recent studies in the literature have highlighted the critical role played by cell signalling in determining haemopoietic stem cell (HSC) fate within ex vivo culture systems. Stimulatory signals can enhance proliferation and promote differentiation, whilst inhibitory signals can significantly limit culture output. METHODS: Numerical models of various mitigation strategies are presented and applied to determine effectiveness of these strategies toward mitigation of paracrine inhibitory signalling inherent in these culture systems. The strategies assessed include mixing, media-exchange, fed-batch and perfusion. RESULTS: The models predict that significant spatial concentration gradients exist in typical cell cultures, with important consequences for subsequent cell expansion. Media exchange is shown to be the most effective mitigation strategy, but remains labour intensive and difficult to scale-up for large culture systems. The fed-batch strategy is only effective at very small Peclet number, and its effect is diminished as the cell culture volume grows. Conversely, mixing is effective at high Peclet number, and ineffective at low Peclet number. The models predict that cell expansion in fed-batch cultures becomes independent of increasing dilution rate, consistent with experimental results previously reported in the literature. In contrast, the models predict that increasing the flow rate in perfused cultures will lead to increased cell expansion, indicating the suitability of perfusion for use as an automated, tunable strategy. The effect of initial cell seeding density is also investigated, with the model showing that perfusion outperforms dilution for all densities considered. CONCLUSIONS: The models predict that the impact of inhibitory signalling in HSC cultures can be mitigated against using media manipulation strategies, with the optimal strategy dependent upon the protein diffusion time-scale relative to the media manipulation time-scale. The key messages from this study can be applied to any complex cell culture scenario where cell-cell interactions and paracrine signalling networks impact upon cell fate and cell expansion. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13287-015-0048-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4443622 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-44436222015-05-27 Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures Berry, Joseph D Godara, Pankaj Liovic, Petar Haylock, David N Stem Cell Res Ther Research INTRODUCTION: Recent studies in the literature have highlighted the critical role played by cell signalling in determining haemopoietic stem cell (HSC) fate within ex vivo culture systems. Stimulatory signals can enhance proliferation and promote differentiation, whilst inhibitory signals can significantly limit culture output. METHODS: Numerical models of various mitigation strategies are presented and applied to determine effectiveness of these strategies toward mitigation of paracrine inhibitory signalling inherent in these culture systems. The strategies assessed include mixing, media-exchange, fed-batch and perfusion. RESULTS: The models predict that significant spatial concentration gradients exist in typical cell cultures, with important consequences for subsequent cell expansion. Media exchange is shown to be the most effective mitigation strategy, but remains labour intensive and difficult to scale-up for large culture systems. The fed-batch strategy is only effective at very small Peclet number, and its effect is diminished as the cell culture volume grows. Conversely, mixing is effective at high Peclet number, and ineffective at low Peclet number. The models predict that cell expansion in fed-batch cultures becomes independent of increasing dilution rate, consistent with experimental results previously reported in the literature. In contrast, the models predict that increasing the flow rate in perfused cultures will lead to increased cell expansion, indicating the suitability of perfusion for use as an automated, tunable strategy. The effect of initial cell seeding density is also investigated, with the model showing that perfusion outperforms dilution for all densities considered. CONCLUSIONS: The models predict that the impact of inhibitory signalling in HSC cultures can be mitigated against using media manipulation strategies, with the optimal strategy dependent upon the protein diffusion time-scale relative to the media manipulation time-scale. The key messages from this study can be applied to any complex cell culture scenario where cell-cell interactions and paracrine signalling networks impact upon cell fate and cell expansion. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13287-015-0048-7) contains supplementary material, which is available to authorized users. BioMed Central 2015-04-16 /pmc/articles/PMC4443622/ /pubmed/25888759 http://dx.doi.org/10.1186/s13287-015-0048-7 Text en © Berry et al.; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Berry, Joseph D Godara, Pankaj Liovic, Petar Haylock, David N Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
title | Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
title_full | Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
title_fullStr | Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
title_full_unstemmed | Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
title_short | Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
title_sort | predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4443622/ https://www.ncbi.nlm.nih.gov/pubmed/25888759 http://dx.doi.org/10.1186/s13287-015-0048-7 |
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