Engineering cells with intracellular agent-loaded microparticles to control cell phenotype

James A Ankrum; Oscar R Miranda; Kelvin S Ng; Debanjan Sarkar; Chenjie Xu; Jeffrey M Karp

doi:10.1038/nprot.2014.002

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Engineering cells with intracellular agent-loaded microparticles to control cell phenotype

Journal article

Peer reviewed

Engineering cells with intracellular agent-loaded microparticles to control cell phenotype

James A Ankrum, Oscar R Miranda, Kelvin S Ng, Debanjan Sarkar, Chenjie Xu and Jeffrey M Karp

Nature protocols, Vol.9(2), pp.233-245

02/2014

DOI: 10.1038/nprot.2014.002

PMCID: PMC4320648

PMID: 24407352

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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.

Phenotype

Particle Size

Cell- and Tissue-Based Therapy - methods

Cell Engineering - methods

Lactic Acid - metabolism

Polyglycolic Acid - metabolism

Drug Delivery Systems - methods

Details

Title: Subtitle: Engineering cells with intracellular agent-loaded microparticles to control cell phenotype
Creators: James A Ankrum - 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA
Oscar R Miranda - 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA
Kelvin S Ng - 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA
Debanjan Sarkar - Department of Biomedical Engineering, University at Buffalo, The State University of New York, Buffalo, New York, USA
Chenjie Xu - Division of Bioengineering, School of Chemical and Biomedical Engineering, Nanyang Technological University, Singapore
Jeffrey M Karp - 1] Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA. Harvard-MIT Division of Health Sciences and Technology, Harvard Stem Cell Institute, Cambridge, Massachusetts, USA
Resource Type: Journal article
Publication Details: Nature protocols, Vol.9(2), pp.233-245
DOI: 10.1038/nprot.2014.002
PMID: 24407352
PMCID: PMC4320648
NLM abbreviation: Nat Protoc
ISSN: 1750-2799
eISSN: 1750-2799
Publisher: England
Grant note: R01 HL095722 / NHLBI NIH HHS HL095722 / NHLBI NIH HHS
Language: English
Date published: 02/2014
Academic Unit: Roy J. Carver Department of Biomedical Engineering
Record Identifier: 9984001077702771

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