title: A microparticle approach to morphogen delivery within pluripotent stem cell aggregates.
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.
authors.affiliation: The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332-0535, USA.
authors.forename: Andrés M
authors.initials: AM
authors.lastname: Bratt-Leal
authors.forename: Anh H
authors.initials: AH
authors.lastname: Nguyen
authors.forename: Katy A
authors.initials: KA
authors.lastname: Hammersmith
authors.forename: Ankur
authors.initials: A
authors.lastname: Singh
authors.forename: Todd C
authors.initials: TC
authors.lastname: McDevitt
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D002449:Cell Aggregation / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D018920:Coculture Techniques; D058732:Embryoid Bodies / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005780:Gelatin / Q000494:pharmacology; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects; D051379:Mice; D008863:Microspheres; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D018874:Spheroids, Cellular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism
chemical_list: D055415:Bone Morphogenetic Protein 4; D005780:Gelatin
keywords: Bone morphogenic protein; Differentiation; Embryoid body; Embryonic stem cells; Mesoderm; Microparticle