title: Engineering cells with intracellular agent-loaded microparticles to control cell phenotype.
abstract: Cell therapies enable unprecedented treatment options to replace tissues, destroy tumors and facilitate regeneration. The greatest challenge facing cell therapy is the inability to control the fate and function of cells after transplantation. We have developed an approach to control cell phenotype in vitro and after transplantation by engineering cells with intracellular depots that continuously release phenotype-altering agents for days to weeks. The platform enables control of cells' secretome, viability, proliferation and differentiation, and the platform can be used to deliver drugs or other factors (e.g., dexamethasone, rhodamine and iron oxide) to the cell's microenvironment. The preparation, efficient internalization and intracellular stabilization of ∼1-μm drug-loaded microparticles are critical for establishing sustained control of cell phenotype. Herein we provide a protocol to generate and characterize micrometer-sized agent-doped poly(lactic-co-glycolic) acid (PLGA) particles by using a single-emulsion evaporation technique (7 h), to uniformly engineer cultured cells (15 h), to confirm particle internalization and to troubleshoot commonly experienced obstacles.
authors.affiliation: 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [2] Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Ankrum
authors.affiliation: 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [2] Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
authors.forename: Oscar R
authors.initials: OR
authors.lastname: Miranda
authors.affiliation: 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [2] Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
authors.forename: Kelvin S
authors.initials: KS
authors.lastname: Ng
authors.affiliation: Department of Biomedical Engineering, University at Buffalo, The State University of New York, Buffalo, New York, USA.
authors.forename: Debanjan
authors.initials: D
authors.lastname: Sarkar
authors.affiliation: Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore.
authors.forename: Chenjie
authors.initials: C
authors.lastname: Xu
authors.affiliation: 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [2] Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Karp
mesh_terms: D060846:Cell Engineering / Q000379:methods*; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D016503:Drug Delivery Systems / Q000379:methods*; D019344:Lactic Acid / Q000378:metabolism*; D010316:Particle Size; D010641:Phenotype; D011100:Polyglycolic Acid / Q000378:metabolism*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer
chemical_list: D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid