Characterization and reduction of non-endocrine cells accompanying islet-like endocrine cells differentiated from human iPSC

  • Sci Rep. 2022 Mar 18;12(1):4740. doi: 10.1038/s41598-022-08753-5.
Hideyuki Hiyoshi   #  1  2 ,  Kensuke Sakuma   #  3  4  5 ,  Noriko Tsubooka-Yamazoe   #  3  4  5 ,  Shinya Asano  6 ,  Taisuke Mochida  3  4 ,  Junji Yamaura  4  7 ,  Shuhei Konagaya  8  4  5 ,  Ryo Fujii  6 ,  Hirokazu Matsumoto  3  4 ,  Ryo Ito  3  4  5 ,  Taro Toyoda  9  10
Affiliations
  • 1. T-CiRA Discovery, Research, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan. [email protected].
  • 2. Takeda-CiRA Joint Program for iPS Cell Applications (T-CiRA), Fujisawa, Kanagawa, Japan. [email protected].
  • 3. T-CiRA Discovery, Research, Takeda Pharmaceutical Company Limited, 26-1, Muraoka-Higashi 2-chome, Fujisawa, Kanagawa, 251-8555, Japan.
  • 4. Takeda-CiRA Joint Program for iPS Cell Applications (T-CiRA), Fujisawa, Kanagawa, Japan.
  • 5. Orizuru Therapeutics, Inc, Fujisawa, Kanagawa, Japan.
  • 6. Axcelead Drug Discovery Partners, Inc, Fujisawa, Kanagawa, Japan.
  • 7. Pharmaceutical Science, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan.
  • 8. Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan.
  • 9. Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. [email protected].
  • 10. Takeda-CiRA Joint Program for iPS Cell Applications (T-CiRA), Fujisawa, Kanagawa, Japan. [email protected].
  • # Contributed equally.
PMID: 35304548 DOI: 10.1038/s41598-022-08753-5
Abstract

The differentiation of pancreatic endocrine cells from human pluripotent stem cells has been thoroughly investigated for their application in cell therapy against diabetes. Although non-endocrine cells are inevitable contaminating by-products of the differentiation process, a comprehensive profile of such cells is lacking. Therefore, we characterized non-endocrine cells in iPSC-derived pancreatic islet cells (iPIC) using single-cell transcriptomic analysis. We found that non-endocrine cells consist of (1) heterogeneous proliferating cells, and (2) cells with not only pancreatic traits but also liver or intestinal traits marked by FGB or AGR2. Non-endocrine cells specifically expressed FGFR2, PLK1, and LDHB. We demonstrated that inhibition of pathways involving these genes selectively reduced the number of non-endocrine cells in the differentiation process. These findings provide useful insights into cell purification approaches and contribute to the improvement of the mass production of endocrine cells for stem cell-derived cell therapy for diabetes.

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