Large-scale production and directed induction of functional dendritic cells ex vivo from serum-free expanded human hematopoietic stem cells

Cytotherapy. 2019 Jul;21(7):755-768. doi: 10.1016/j.jcyt.2019.04.059. Epub 2019 May 17.

Abstract

Background: Dendritic cells (DCs) that are derived from hematopoietic stem cells (HSCs) are the most potent antigen-presenting cells and play a pivotal role in initiating the immune response. Hence, large-scale production and direct induction of functional DCs ex vivo from HSCs are crucial to HSC research and clinical potential, such as vaccines for cancer and immune therapy.

Methods: In a previous study, we developed a serum-free HSC expansion system (SF-HSC medium) to expand large numbers of primitive HSCs ex vivo. Herein, a DC induction and expansion medium (DC medium) was proposed to further generate large numbers of functional DCs from serum-free expanded HSCs, which were developed and optimized by factorial design and the steepest ascent method.

Results: The DC medium is composed of effective basal medium (Iscove's modified Dulbecco's medium [IMDM]) and cytokines (2.9 ng/mL stem cell factor [SCF], 2.1 ng/mL Flt-3 ligand, 3.6 ng/mL interleukin [IL]-1β, 19.3 ng/mL granulocyte-macrophage colony-stimulating factor [GM-CSF] and 20.0 ng/mL tumor necrosis factor-α [TNF-α]). After 10-day culture in DC medium, the maximum fold expansion for accumulated CD1a+CD11c+ DCs was more than 4000-fold, and the induced DCs were characterized and confirmed by analysis of growth kinetics, surface antigen expression, endocytosis ability, mixed lymphocyte reaction, specific cytokine secretion and lipopolysaccharide stimulation.

Discussion: In conclusion, the combination of DC medium and SF-HSC medium can efficiently induce and expand a large amount of functional DCs from a small scale of HSCs and might be a promising source of DCs for vaccine and immune therapy in the near future.

Keywords: dendritic cells; ex vivo expansion; hematopoietic stem cells; immunotherapy; induction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD34 / metabolism
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Culture Media, Serum-Free / pharmacology*
  • Cytokines / metabolism
  • Dendritic Cells / cytology*
  • Dendritic Cells / drug effects
  • Dendritic Cells / physiology
  • Endocytosis
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / physiology
  • Humans
  • Lipopolysaccharides / pharmacology
  • Lymphocyte Culture Test, Mixed
  • Stem Cell Factor / pharmacology

Substances

  • Antigens, CD34
  • Culture Media, Serum-Free
  • Cytokines
  • Lipopolysaccharides
  • Stem Cell Factor