A microparticle approach to morphogen delivery within pluripotent stem cell aggregates

• Type of Material: Article
• Authors: Andres M. Bratt-Leal, Georgia Institute of Technology; Anh H. Nguyen, Georgia Institute of Technology; Katy A. Hammersmith, Georgia Institute of Technology; Ankur Singh, Georgia Institute of Technology; Todd McDevitt, Emory University
• Language: English
• Date: 2013-10-01
• Publisher: Elsevier
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2013 Elsevier Ltd. All rights reserved
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.biomaterials.2013.05.079
• Title of Journal or Parent Work: Biomaterials
• Volume: 34
• Issue: 30
• Start Page: 7227
• End Page: 7235
• Grant/Funding Information: Financial support was provided by funding from the National Institutes of Health (GM088291) and the National Science Foundation (CBET 0651739).; AN was supported by an NSF Graduate Research Fellowship.; ABL was supported by an NIH training grant (GM008433) as well as funding from the Goizueta Foundation.
• Keywords: Differentiation; Progenitor cells; Specification; Embryoid body; Heparan sulfate; Materials Science, Biomaterials; Materials Science; Bodies; Embryoid body differentiation; Mesoderm; Technology; Science & Technology; Endothelial growth factor; Embryonic stem cells; VEGF; Microparticle; Bone morphogenic protein; Engineering, Biomedical; Organization; Protein 4; Engineering
• Research Categories: Biology, Cell; Engineering, Biomedical; Engineering, Mechanical; Biology, Microbiology

Abstract

Stem cell fate and specification is largely controlled by extrinsic cues that comprise the 3D microenvironment. Biomaterials can serve to control the spatial and temporal presentation of morphogenic molecules in order to direct stem cell fate decisions. Here we describe a microparticle (MP)-based approach to deliver growth factors within multicellular aggregates to direct pluripotent stem cell differentiation. Compared to conventional soluble delivery methods, gelatin MPs laden with BMP4 or noggin induced efficient gene expression of mesoderm and ectoderm lineages, respectively, despite using nearly 12-fold less total growth factor. BMP4-laden MPs increased the percentage of cells expressing GFP under the control of the Brachyury-T promoter as visualized by whole-mount confocal imaging and quantified by flow cytometry. Furthermore, the ability to localize MPs laden with different morphogens within a particular hemisphere of stem cell aggregates allowed for spatial control of differentiation within 3D cultures. Overall, localized delivery of growth factors within multicellular aggregates from microparticle delivery vehicles is an important step towards scalable differentiation technologies and the study of morphogen gradients in pluripotent stem cell differentiation.