Differentiation of Human-Induced Pluripotent Stem Cell-Derived Endocrine Progenitors to Islet-like Cells Using a Dialysis Suspension Culture System

Cells. 2021 Aug 7;10(8):2017. doi: 10.3390/cells10082017.

Abstract

The production of functional islet-like cells from human-induced pluripotent stem cells (hiPSCs) is a promising strategy for the therapeutic use and disease modeling for type 1 diabetes. However, the production cost of islet-like cells is extremely high due to the use of expensive growth factors for differentiation. In a conventional culture method, growth factors and beneficial autocrine factors remaining in the culture medium are removed along with toxic metabolites during the medium change, and it limits the efficient utilization of those factors. In this study, we demonstrated that the dialysis suspension culture system is possible to reduce the usage of growth factors to one-third in the differentiation of hiPSC-derived endocrine progenitor cells to islet-like cells by reducing the medium change frequency with the refinement of the culture medium. Furthermore, the expression levels of hormone-secretion-related genes and the efficiency of differentiation were improved with the dialysis suspension culture system, possibly due to the retaining of autocrine factors. In addition, we confirmed several improvements required for the further study of the dialysis culture system. These findings showed the promising possibility of the dialysis suspension culture system for the low-cost production of islet-like cells.

Keywords: cost reduction; dialysis; differentiation; hiPSC; islet-like cell; pancreatic β cell; suspension culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Cell Aggregation / drug effects
  • Cell Aggregation / genetics
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects*
  • Cell Differentiation / genetics
  • Cell Line
  • Culture Media / chemistry
  • Culture Media / pharmacology*
  • Dialysis Solutions / chemistry
  • Dialysis Solutions / pharmacology*
  • Endocrine System / cytology
  • Endocrine System / drug effects
  • Endocrine System / metabolism
  • Gene Expression / drug effects
  • Glucose Transporter Type 2 / genetics
  • Glucose Transporter Type 2 / metabolism
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects*
  • Induced Pluripotent Stem Cells / metabolism
  • Islets of Langerhans / cytology
  • Islets of Langerhans / drug effects*
  • Islets of Langerhans / metabolism
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • Renal Dialysis*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Trans-Activators / genetics
  • Trans-Activators / metabolism

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Culture Media
  • Dialysis Solutions
  • Glucose Transporter Type 2
  • Homeodomain Proteins
  • NEUROG3 protein, human
  • Nerve Tissue Proteins
  • Trans-Activators
  • pancreatic and duodenal homeobox 1 protein