A Novel High-Throughput Screening Platform Reveals an Optimized Cytokine Formulation for Human Hematopoietic Progenitor Cell Expansion

Stem Cells Dev. 2016 Nov 15;25(22):1709-1720. doi: 10.1089/scd.2016.0216. Epub 2016 Sep 30.


The main limitations of hematopoietic cord blood (CB) transplantation, viz, low cell dosage and delayed reconstitution, can be overcome by ex vivo expansion. CB expansion under conventional culture causes rapid cell differentiation and depletion of hematopoietic stem and progenitor cells (HSPCs) responsible for engraftment. In this study, we use combinatorial cell culture technology (CombiCult®) to identify medium formulations that promote CD133+ CB HSPC proliferation while maintaining their phenotypic characteristics. We employed second-generation CombiCult screens that use electrospraying technology to encapsulate CB cells in alginate beads. Our results suggest that not only the combination but also the order of addition of individual components has a profound influence on expansion of specific HSPC populations. Top protocols identified by the CombiCult screen were used to culture human CD133+ CB HSPCs on nanofiber scaffolds and validate the expansion of the phenotypically defined CD34+CD38lo/-CD45RA-CD90+CD49f+ population of hematopoietic stem cells and their differentiation into defined progeny.

Keywords: CombiCult; alginate encapsulation; ex-vivo expansion; hematopoietic stem/progenitor cells; high-throughput-screening.

MeSH terms

  • AC133 Antigen / metabolism
  • Alginates / pharmacology
  • Algorithms
  • Cell Adhesion / drug effects
  • Cell Proliferation / drug effects
  • Cytokines / pharmacology*
  • Flow Cytometry
  • Glucuronic Acid / pharmacology
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism
  • Hexuronic Acids / pharmacology
  • High-Throughput Screening Assays / methods*
  • Humans
  • Microspheres
  • Nanofibers / chemistry
  • Reproducibility of Results
  • Tissue Scaffolds / chemistry


  • AC133 Antigen
  • Alginates
  • Cytokines
  • Hexuronic Acids
  • Glucuronic Acid