title: Spatiotemporal proliferation of human stromal cells adjusts to nutrient availability and leads to stanniocalcin-1 expression in vitro and in vivo.
abstract: Cells and tissues are intrinsically adapted to molecular gradients and use them to maintain or change their activity. The effect of such gradients is particularly important for cell populations that have an intrinsic capacity to differentiate into multiple cell lineages, such as bone marrow derived mesenchymal stromal cells (MSCs). Our results showed that nutrient gradients prompt the spatiotemporal organization of MSCs in 3D culture. Cells adapted to their 3D environment without significant cell death or cell differentiation. Kinetics data and whole-genome gene expression analysis suggest that a low proliferation activity phenotype predominates in stromal cells cultured in 3D, likely due to increasing nutrient limitation. These differences implied that despite similar surface areas available for cell attachment, higher cell concentrations in 3D reduced MSCs proliferation, while activating hypoxia related-pathways. To further understand the in vivo effects of both proliferation and cell concentrations, we increased cell concentrations in small (1.8 μl) implantable wells. We found that MSCs accumulation and conditioning by nutrient competition in small volumes leads to an ideal threshold of cell-concentration for the induction of blood vessel formation, possibly signaled by the hypoxia-related stanniocalcin-1 gene.
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands. Electronic address: g.higuerasierra@erasmusmc.nl.
authors.forename: Gustavo A
authors.initials: GA
authors.lastname: Higuera
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Hugo
authors.initials: H
authors.lastname: Fernandes
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Tim W G M
authors.initials: TW
authors.lastname: Spitters
authors.affiliation: Erasmus Medical Center, Internal Medicine, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands.
authors.forename: Jeroen
authors.initials: J
authors.lastname: van de Peppel
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Nils
authors.initials: N
authors.lastname: Aufferman
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Roman
authors.initials: R
authors.lastname: Truckenmueller
authors.affiliation: Biophysical Engineering Group, Mesa(+) Institute for Nanotechnology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Maryana
authors.initials: M
authors.lastname: Escalante
authors.affiliation: TNO, Metabolic Health Research, Zernikedreef 9, 2333 CK Leiden, The Netherlands.
authors.forename: Reinout
authors.initials: R
authors.lastname: Stoop
authors.affiliation: Erasmus Medical Center, Internal Medicine, Wytemaweg 80, 3015 CN Rotterdam, The Netherlands.
authors.forename: Johannes P
authors.initials: JP
authors.lastname: van Leeuwen
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: de Boer
authors.affiliation: Biophysical Engineering Group, Mesa(+) Institute for Nanotechnology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Vinod
authors.initials: V
authors.lastname: Subramaniam
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Karperien
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Clemens
authors.initials: C
authors.lastname: van Blitterswijk
authors.affiliation: Systems and Control Group, Wageningen University, PO Box 17, 6700 AA Wageningen, The Netherlands.
authors.forename: Anton
authors.initials: A
authors.lastname: van Boxtel
authors.affiliation: Department of Tissue Regeneration, MIRA - Institute for Biomedical Technology, University of Twente, Drienerlolaan 5, 7522 NB Enschede, The Netherlands.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Moroni
mesh_terms: D000222:Adaptation, Physiological / Q000502:physiology; D061249:Batch Cell Culture Techniques / Q000379:methods*; D001682:Biological Availability; D049109:Cell Proliferation / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000493:pharmacokinetics*; D004867:Equipment Design; D006023:Glycoproteins / Q000096:biosynthesis*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D018919:Neovascularization, Physiologic / Q000502:physiology*; D062211:Spatio-Temporal Analysis; D054457:Tissue Scaffolds
chemical_list: D003470:Culture Media; D006023:Glycoproteins; C051795:teleocalcin
keywords: Angiogenesis; ECM (extracellular matrix); Mesenchymal stromal cells; Microarchitecture; Molecular gradients