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

Nat Protoc. 2014 Feb;9(2):233-45. doi: 10.1038/nprot.2014.002. Epub 2014 Jan 9.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Engineering / methods*
  • Cell- and Tissue-Based Therapy / methods*
  • Drug Delivery Systems / methods*
  • Lactic Acid / metabolism*
  • Particle Size
  • Phenotype*
  • Polyglycolic Acid / metabolism*
  • Polylactic Acid-Polyglycolic Acid Copolymer


  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polyglycolic Acid
  • Lactic Acid