Short-range cytokine gradients to mimic paracrine cell interactions in vitro

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• 作者：Michael Ansorge^a ; Nadine Rastig^a ; Ralph Steinborn^a ; Tina Kö ; nig^a ; Lars Baumann^a ; Stephanie Mö ; ller^b ; Matthias Schnabelrauch^b ; Michael Cross^c ; Carsten Werner^d ; ^e ; Annette G. Beck-Sickinger^a ; Tilo Pompe^a ; ^d ; ^{tilo.pompe@uni-leipzig.de" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Gradients ; Chemokine ; Chemotaxis ; Agarose ; Glycosaminoglycan ; Microparticle ; Affinity-based release
• 刊名：Journal of Controlled Release
• 出版年：2016
• 出版时间：28 February 2016
• 年：2016
• 卷：224
• 期：Complete
• 页码：59-68
• 全文大小：1047 K

文摘

Cell fate decisions in many physiological processes, including embryogenesis, stem cell niche homeostasis and wound healing, are regulated by secretion of small signaling proteins, called cytokines, from source cells to their neighbors or into the environment. Concentration level and steepness of the resulting paracrine gradients elicit different cell responses, including proliferation, differentiation or chemotaxis. For an in-depth analysis of underlying mechanisms, in vitro models are required to mimic in vivo cytokine gradients. We set up a microparticle-based system to establish short-range cytokine gradients in a three-dimensional extracellular matrix context. To provide native binding sites for cytokines, agarose microparticles were functionalized with different glycosaminoglycans (GAG). After protein was loaded onto microparticles, its slow release was quantified by confocal microscopy and fluorescence correlation spectroscopy. Besides the model protein lysozyme, SDF-1 was used as a relevant chemokine for hematopoietic stem and progenitor cell (HSPC) chemotaxis. For both proteins we found gradients ranging up to 50 μm from the microparticle surface and concentrations in the order of nM to pM in dependence on loading concentration and affinity modulation by the GAG functionalization. Directed chemotactic migration of cells from a hematopoietic cell line (FDCPmix) and primary murine HSPC (Sca-1⁺ CD150⁺ CD48⁻) toward the SDF-1-laden microparticles proved functional short-range gradients in a two-dimensional and three-dimensional setting over time periods of many hours. The approach has the potential to be applied to other cytokines mimicking paracrine cell–cell interactions in vitro.