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SLAS Technol. 2021 Jun;26(3):287-299. doi: 10.1177/2472630320972110. Epub 2020 Dec 09.

Automating Human Induced Pluripotent Stem Cell Culture and Differentiation of iPSC-Derived Retinal Pigment Epithelium for Personalized Drug Testing.

SLAS technology

Vincent Truong, Kevin Viken, Zhaohui Geng, Samantha Barkan, Blake Johnson, Mara C Ebeling, Sandra R Montezuma, Deborah A Ferrington, James R Dutton

Affiliations

  1. Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA.
  2. Department of Ophthalmology and Visual Neurosciences, University of Minnesota, Minneapolis, MN, USA.
  3. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA.

PMID: 33292045 PMCID: PMC8140989 DOI: 10.1177/2472630320972110

Abstract

Derivation and differentiation of human induced pluripotent stem cells (hiPSCs) provide the opportunity to generate medically important cell types from individual patients and patient populations for research and the development of potential cell therapies. This technology allows disease modeling and drug screening to be carried out using diverse population cohorts and with more relevant cell phenotypes than can be accommodated using traditional immortalized cell lines. However, technical complexities in the culture and differentiation of hiPSCs, including lack of scale and standardization and prolonged experimental timelines, limit the adoption of this technology for many large-scale studies, including personalized drug screening. The entry of reproducible end-to-end automated workflows for hiPSC culture and differentiation, demonstrated on commercially available platforms, provides enhanced accessibility of this technology for both research laboratories and commercial pharmaceutical testing. Here we have utilized TECAN Fluent automated cell culture workstations to perform hiPSC culture and differentiation in a reproducible and scalable process to generate patient-derived retinal pigment epithelial cells for downstream use, including drug testing. hiPSCs derived from multiple donors with age-related macular degeneration (AMD) were introduced into our automated workflow, and cell lines were cultured and differentiated into retinal pigment epithelium (RPE). Donor hiPSC-RPE lines were subsequently entered in an automated drug testing workflow to measure mitochondrial function after exposure to "mitoactive" compounds. This work demonstrates scalable, reproducible culture and differentiation of hiPSC lines from individuals on the TECAN Fluent platform and illustrates the potential for end-to-end automation of hiPSC-based personalized drug testing.

Keywords: TECAN Fluent; age-related macular degeneration; automated cell culture platform; automation; human induced pluripotent stem cells; liquid handling; personalized drug screening; retinal pigment epithelium

References

  1. Front Aging Neurosci. 2014 Aug 01;6:191 - PubMed
  2. PLoS One. 2013;8(1):e53764 - PubMed
  3. Stem Cells Transl Med. 2015 Mar;4(3):217-23 - PubMed
  4. Nature. 2008 Jan 10;451(7175):141-6 - PubMed
  5. Sci Rep. 2019 Jul 23;9(1):10646 - PubMed
  6. SLAS Discov. 2017 Sep;22(8):1016-1025 - PubMed
  7. Pharmacol Rev. 2020 Jan;72(1):320-342 - PubMed
  8. Nat Rev Drug Discov. 2017 Feb;16(2):115-130 - PubMed
  9. Exp Eye Res. 2019 Aug;185:107641 - PubMed
  10. N Engl J Med. 2017 Mar 16;376(11):1038-1046 - PubMed
  11. Stem Cells Transl Med. 2013 May;2(5):384-93 - PubMed
  12. Methods Mol Biol. 2016;1357:221-9 - PubMed
  13. J Am Coll Cardiol. 2016 May 10;67(18):2161-2176 - PubMed
  14. Physiol Rev. 2005 Jul;85(3):845-81 - PubMed
  15. Redox Biol. 2017 Oct;13:255-265 - PubMed
  16. PLoS One. 2017 Mar 10;12(3):e0173575 - PubMed
  17. Cell Stem Cell. 2017 May 4;20(5):635-647.e7 - PubMed
  18. Cell. 2007 Nov 30;131(5):861-72 - PubMed
  19. Stem Cells Transl Med. 2015 Apr;4(4):381-8 - PubMed
  20. Stem Cells Transl Med. 2019 May;8(5):466-477 - PubMed
  21. Redox Biol. 2020 Jul;34:101552 - PubMed
  22. Clin Ophthalmol. 2008 Jun;2(2):413-24 - PubMed
  23. Nat Rev Genet. 2019 Jul;20(7):377-388 - PubMed
  24. J Cell Sci Suppl. 1993;17:189-95 - PubMed
  25. Lancet. 2015 Feb 7;385(9967):509-16 - PubMed
  26. Ophthalmol Retina. 2020 Mar;4(3):264-273 - PubMed
  27. Stem Cells. 2017 Nov;35(11):2305-2320 - PubMed
  28. Invest Ophthalmol Vis Sci. 2004 Dec;45(12):4484-90 - PubMed

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