Microfluidic perfusion for regulating diffusible signaling in stem cells.

<h4>Background</h4>Autocrine & paracrine signaling are widespread both in vivo and in vitro, and are particularly important in embryonic stem cell (ESC) pluripotency and lineage commitment. Although autocrine signaling via fibroblast growth factor-4 (FGF4) is known to be required in...

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Autores principales:	Katarina Blagovic, Lily Y Kim, Joel Voldman
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ef28389e825a42d8913d5b8480e7b206
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spelling	oai:doaj.org-article:ef28389e825a42d8913d5b8480e7b2062021-11-18T06:48:39ZMicrofluidic perfusion for regulating diffusible signaling in stem cells.1932-620310.1371/journal.pone.0022892https://doaj.org/article/ef28389e825a42d8913d5b8480e7b2062011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21829665/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Autocrine & paracrine signaling are widespread both in vivo and in vitro, and are particularly important in embryonic stem cell (ESC) pluripotency and lineage commitment. Although autocrine signaling via fibroblast growth factor-4 (FGF4) is known to be required in mouse ESC (mESC) neuroectodermal specification, the question of whether FGF4 autocrine signaling is sufficient, or whether other soluble ligands are also involved in fate specification, is unknown. The spatially confined and closed-loop nature of diffusible signaling makes its experimental control challenging; current experimental approaches typically require prior knowledge of the factor/receptor in order to modulate the loop. A new approach explored in this work is to leverage transport phenomena at cellular resolution to downregulate overall diffusible signaling through the physical removal of cell-secreted ligands.<h4>Methodology/principal findings</h4>We develop a multiplex microfluidic platform to continuously remove cell-secreted (autocrine\paracrine) factors to downregulate diffusible signaling. By comparing cell growth and differentiation in side-by-side chambers with or without added cell-secreted factors, we isolate the effects of diffusible signaling from artifacts such as shear, nutrient depletion, and microsystem effects, and find that cell-secreted growth factor(s) are required during neuroectodermal specification. Then we induce FGF4 signaling in minimal chemically defined medium (N2B27) and inhibit FGF signaling in fully supplemented differentiation medium with cell-secreted factors to determine that the non-FGF cell-secreted factors are required to promote growth of differentiating mESCs.<h4>Conclusions/significance</h4>Our results demonstrate for the first time that flow can downregulate autocrine\paracrine signaling and examine sufficiency of extracellular factors. We show that autocrine\paracrine signaling drives neuroectodermal commitment of mESCs through both FGF4-dependent and -independent pathways. Overall, by uncovering autocrine\paracrine processes previously hidden in conventional culture systems, our results establish microfluidic perfusion as a technique to study and manipulate diffusible signaling in cell systems.Katarina BlagovicLily Y KimJoel VoldmanPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 8, p e22892 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Katarina Blagovic Lily Y Kim Joel Voldman Microfluidic perfusion for regulating diffusible signaling in stem cells.
description	<h4>Background</h4>Autocrine & paracrine signaling are widespread both in vivo and in vitro, and are particularly important in embryonic stem cell (ESC) pluripotency and lineage commitment. Although autocrine signaling via fibroblast growth factor-4 (FGF4) is known to be required in mouse ESC (mESC) neuroectodermal specification, the question of whether FGF4 autocrine signaling is sufficient, or whether other soluble ligands are also involved in fate specification, is unknown. The spatially confined and closed-loop nature of diffusible signaling makes its experimental control challenging; current experimental approaches typically require prior knowledge of the factor/receptor in order to modulate the loop. A new approach explored in this work is to leverage transport phenomena at cellular resolution to downregulate overall diffusible signaling through the physical removal of cell-secreted ligands.<h4>Methodology/principal findings</h4>We develop a multiplex microfluidic platform to continuously remove cell-secreted (autocrine\paracrine) factors to downregulate diffusible signaling. By comparing cell growth and differentiation in side-by-side chambers with or without added cell-secreted factors, we isolate the effects of diffusible signaling from artifacts such as shear, nutrient depletion, and microsystem effects, and find that cell-secreted growth factor(s) are required during neuroectodermal specification. Then we induce FGF4 signaling in minimal chemically defined medium (N2B27) and inhibit FGF signaling in fully supplemented differentiation medium with cell-secreted factors to determine that the non-FGF cell-secreted factors are required to promote growth of differentiating mESCs.<h4>Conclusions/significance</h4>Our results demonstrate for the first time that flow can downregulate autocrine\paracrine signaling and examine sufficiency of extracellular factors. We show that autocrine\paracrine signaling drives neuroectodermal commitment of mESCs through both FGF4-dependent and -independent pathways. Overall, by uncovering autocrine\paracrine processes previously hidden in conventional culture systems, our results establish microfluidic perfusion as a technique to study and manipulate diffusible signaling in cell systems.
format	article
author	Katarina Blagovic Lily Y Kim Joel Voldman
author_facet	Katarina Blagovic Lily Y Kim Joel Voldman
author_sort	Katarina Blagovic
title	Microfluidic perfusion for regulating diffusible signaling in stem cells.
title_short	Microfluidic perfusion for regulating diffusible signaling in stem cells.
title_full	Microfluidic perfusion for regulating diffusible signaling in stem cells.
title_fullStr	Microfluidic perfusion for regulating diffusible signaling in stem cells.
title_full_unstemmed	Microfluidic perfusion for regulating diffusible signaling in stem cells.
title_sort	microfluidic perfusion for regulating diffusible signaling in stem cells.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/ef28389e825a42d8913d5b8480e7b206
work_keys_str_mv	AT katarinablagovic microfluidicperfusionforregulatingdiffusiblesignalinginstemcells AT lilyykim microfluidicperfusionforregulatingdiffusiblesignalinginstemcells AT joelvoldman microfluidicperfusionforregulatingdiffusiblesignalinginstemcells
_version_	1718424389883527168

Microfluidic perfusion for regulating diffusible signaling in stem cells.

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