Fabricating retinal pigment epithelial cell sheets derived from human induced pluripotent stem cells in an automated closed culture system for regenerative medicine

PLoS One. 2019 Mar 13;14(3):e0212369. doi: 10.1371/journal.pone.0212369. eCollection 2019.

Abstract

Regenerative medicine has received a lot of attention as a novel strategy for injuries and diseases that are difficult to cure using current techniques. Cell production, which is vital for regenerative medicine, has undergone remarkable progress via breakthroughs in developmental biology and tissue engineering; currently, cell production requires numerous experimental operators performing manual, small-scale cell cultures. Other major obstacles for cell production and regenerative medicine include the variable quality of products based on the experimental procedure, the skills of operators, the level of labor required for production, and costs. Technological developments are required to overcome this, including automation instead of manual culture. Age-related macular regeneration (AMD) is a refractory ocular disease that causes severe deterioration in central vision due to senescence in the retinal pigment epithelium (RPE). Recently, we performed an autologous transplantation of induced pluripotent stem (iPS) cell-derived RPE cell sheets and started clinical research on allografts from RPE cell suspensions differentiated from iPS cells. The use of regenerative therapies for AMD using iPS cell-derived RPE is expected to become more widespread. In the present study, human iPS cell-derived RPE cells were cultured to form RPE cell sheets using equipment with a closed culture module. The quality of the automated cultured RPE cell sheets was confirmed by comparing their morphological and biological properties with those of manually generated RPE cell sheets. As a result, machine-cultured RPE sheets displayed the same quality as manually cultured RPE sheets, showing that iPS cell-derived RPE cell sheets were successfully cultured by an automated process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation, Laboratory
  • Cell Culture Techniques / methods
  • Cells, Cultured
  • Eye Proteins / metabolism
  • Feasibility Studies
  • Humans
  • Immunohistochemistry
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Macular Degeneration / therapy
  • Nerve Growth Factors / metabolism
  • Regenerative Medicine / methods*
  • Retinal Pigment Epithelium / cytology*
  • Retinal Pigment Epithelium / metabolism
  • Serpins / metabolism
  • Tissue Engineering / methods
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Eye Proteins
  • Nerve Growth Factors
  • Serpins
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • pigment epithelium-derived factor

Grants and funding

Development of ACE3, we used in this study, was partly supported by research grants "Creation of Innovation Centers for Advanced Interdisciplinary Research Areas Program" from Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (to S.T. and H.H.). A part of this work was supported by internal funds from Hitachi Research & Development Group to RIKEN Center for Biosystems Dynamics Research (to M.T. and N.K.) Hitachi and Sanplatec are commercial companies. They provided financial support in the form of authors’ salaries and research materials for the authors (E.M., H.H., M.K., M.O., J.K., and S.T.) during the conduct of the study. However, these companies did not have any additional roles in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.