A Technique for Removing Tumourigenic Pluripotent Stem Cells Using RBC2LCN Lectin

Overview

Journal Regen Ther

Date 2020 May 29

PMID 32462059

Citations 6

Authors

Yoshikazu Haramoto

Yasuko Onuma

Shuuji Mawaribuchi

Yoshiro Nakajima

Yasuhiko Aiki

Kumiko Higuchi

Madoka Shimizu

Hiroaki Tateno

Jun Hirabayashi

Yuzuru Ito

Affiliations

Soon will be listed here.

Abstract

Introduction: Tumourigenesis attributed to residual undifferentiated cells in a graft is considered to be a significant issue in cell therapy using human pluripotent stem cells. To ensure the safety of regenerative medicine derived from pluripotent stem cells, residual undifferentiated cells must be eliminated in the manufacturing process. We previously described the lectin probe rBC2LCN, which binds harmlessly and specifically to the cell surface of human pluripotent stem cells. We report here a technique using rBC2LCN to remove pluripotent cells from a heterogenous population to reduce the chance of teratoma formation.

Methods: We demonstrate a method for separating residual tumourigenic cells using rBC2LCN-bound magnetic beads. This technology is a novel use of their previous discovery that rBC2LCN is a lectin that selectively binds to pluripotent cells. We optimize and validate a method to remove hPSCs from a mixture with human fibroblasts using rBC2LCN-conjugated magnetic beads.

Results: Cells with the potential to form teratoma could be effectively eliminated from a heterogeneous cell population with biotin-labelled rBC2LCN and streptavidin-bound magnetic beads. The efficiency was measured by FACS, ddPCR, and animal transplantation, suggesting that magnetic cell separation using rBC2LCN is quite efficient for eliminating hPSCs from mixed cell populations.

Conclusions: The removal of residual tumourigenic cells based on rBC2LCN could be a practical option for laboratory use and industrialisation of regenerative medicine using human pluripotent stem cells.

Citing Articles

Label-Free and High-Throughput Removal of Residual Undifferentiated Cells From iPSC-Derived Spinal Cord Progenitor Cells.

Nguyen T, Chooi W, Jeon H, Chen J, Tan J, Roxby D Stem Cells Transl Med. 2024; 13(4):387-398.

PMID: 38321361 PMC: 11016845. DOI: 10.1093/stcltm/szae002.

BiotechLec: an interactive guide of commercial lectins for glycobiology and biomedical research applications.

Schnider B, Escudero F, Imberty A, Lisacek F Glycobiology. 2023; 33(9):684-686.

PMID: 37083961 PMC: 10627245. DOI: 10.1093/glycob/cwad034.

Methods to Assess Proliferation of Stimulated Human Lymphocytes In Vitro: A Narrative Review.

Ganesan N, Ronsmans S, Hoet P Cells. 2023; 12(3).

PMID: 36766728 PMC: 9913443. DOI: 10.3390/cells12030386.

Inhibition of Epithelial-Mesenchymal Transition Maintains Stemness in Human Amniotic Epithelial Cells.

Takano C, Horie M, Taiko I, Trinh Q, Kanemaru K, Komine-Aizawa S Stem Cell Rev Rep. 2022; 18(8):3083-3091.

PMID: 35931939 PMC: 9622541. DOI: 10.1007/s12015-022-10420-1.

CellTrace™ Violet Flow Cytometric Assay to Assess Cell Proliferation.

Lemieszek M, Findlay S, Siegers G Methods Mol Biol. 2022; 2508:101-114.

PMID: 35737236 DOI: 10.1007/978-1-0716-2376-3_9.

References

Roy N, Cleren C, Singh S, Yang L, Beal M, Goldman S . Functional engraftment of human ES cell-derived dopaminergic neurons enriched by coculture with telomerase-immortalized midbrain astrocytes. Nat Med. 2006; 12(11):1259-68. DOI: 10.1038/nm1495. View

Hentze H, Soong P, Wang S, Phillips B, Putti T, Dunn N . Teratoma formation by human embryonic stem cells: evaluation of essential parameters for future safety studies. Stem Cell Res. 2009; 2(3):198-210. DOI: 10.1016/j.scr.2009.02.002. View

Przyborski S . Differentiation of human embryonic stem cells after transplantation in immune-deficient mice. Stem Cells. 2005; 23(9):1242-50. DOI: 10.1634/stemcells.2005-0014. View

Ben-David U, Gan Q, Golan-Lev T, Arora P, Yanuka O, Oren Y . Selective elimination of human pluripotent stem cells by an oleate synthesis inhibitor discovered in a high-throughput screen. Cell Stem Cell. 2013; 12(2):167-79. DOI: 10.1016/j.stem.2012.11.015. View

Kikuchi T, Morizane A, Doi D, Magotani H, Onoe H, Hayashi T . Human iPS cell-derived dopaminergic neurons function in a primate Parkinson's disease model. Nature. 2017; 548(7669):592-596. DOI: 10.1038/nature23664. View

Prokhorova T, Harkness L, Frandsen U, Ditzel N, Schroder H, Burns J . Teratoma formation by human embryonic stem cells is site dependent and enhanced by the presence of Matrigel. Stem Cells Dev. 2008; 18(1):47-54. DOI: 10.1089/scd.2007.0266. View

Choo A, Tan H, Ang S, Fong W, Chin A, Lo J . Selection against undifferentiated human embryonic stem cells by a cytotoxic antibody recognizing podocalyxin-like protein-1. Stem Cells. 2008; 26(6):1454-63. DOI: 10.1634/stemcells.2007-0576. View

Kawamura M, Miyagawa S, Fukushima S, Saito A, Miki K, Ito E . Enhanced survival of transplanted human induced pluripotent stem cell-derived cardiomyocytes by the combination of cell sheets with the pedicled omental flap technique in a porcine heart. Circulation. 2013; 128(11 Suppl 1):S87-94. DOI: 10.1161/CIRCULATIONAHA.112.000366. View

Sulak O, Cioci G, Delia M, Lahmann M, Varrot A, Imberty A . A TNF-like trimeric lectin domain from Burkholderia cenocepacia with specificity for fucosylated human histo-blood group antigens. Structure. 2010; 18(1):59-72. DOI: 10.1016/j.str.2009.10.021. View

10.

Kuroda T, Yasuda S, Matsuyama S, Tano K, Kusakawa S, Sawa Y . Highly sensitive droplet digital PCR method for detection of residual undifferentiated cells in cardiomyocytes derived from human pluripotent stem cells. Regen Ther. 2019; 2:17-23. PMC: 6581767. DOI: 10.1016/j.reth.2015.08.001. View

11.

Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K . Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007; 131(5):861-72. DOI: 10.1016/j.cell.2007.11.019. View

12.

Andrews P, Banting G, Damjanov I, Arnaud D, Avner P . Three monoclonal antibodies defining distinct differentiation antigens associated with different high molecular weight polypeptides on the surface of human embryonal carcinoma cells. Hybridoma. 1984; 3(4):347-61. DOI: 10.1089/hyb.1984.3.347. View

13.

Wang Y, Nakagawa M, Garitaonandia I, Slavin I, Altun G, Lacharite R . Specific lectin biomarkers for isolation of human pluripotent stem cells identified through array-based glycomic analysis. Cell Res. 2011; 21(11):1551-63. PMC: 3364725. DOI: 10.1038/cr.2011.148. View

14.

Tang C, Lee A, Volkmer J, Sahoo D, Nag D, Mosley A . An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells. Nat Biotechnol. 2011; 29(9):829-34. PMC: 3537836. DOI: 10.1038/nbt.1947. View

15.

Tateno H, Onuma Y, Ito Y, Minoshima F, Saito S, Shimizu M . Elimination of tumorigenic human pluripotent stem cells by a recombinant lectin-toxin fusion protein. Stem Cell Reports. 2015; 4(5):811-20. PMC: 4437484. DOI: 10.1016/j.stemcr.2015.02.016. View

16.

Tateno H, Toyota M, Saito S, Onuma Y, Ito Y, Hiemori K . Glycome diagnosis of human induced pluripotent stem cells using lectin microarray. J Biol Chem. 2011; 286(23):20345-53. PMC: 3121447. DOI: 10.1074/jbc.M111.231274. View

17.

Parr C, Katayama S, Miki K, Kuang Y, Yoshida Y, Morizane A . MicroRNA-302 switch to identify and eliminate undifferentiated human pluripotent stem cells. Sci Rep. 2016; 6:32532. PMC: 5016789. DOI: 10.1038/srep32532. View

18.

Thomson J, Itskovitz-Eldor J, Shapiro S, Waknitz M, Swiergiel J, Marshall V . Embryonic stem cell lines derived from human blastocysts. Science. 1998; 282(5391):1145-7. DOI: 10.1126/science.282.5391.1145. View

19.

Ben-David U, Benvenisty N . The tumorigenicity of human embryonic and induced pluripotent stem cells. Nat Rev Cancer. 2011; 11(4):268-77. DOI: 10.1038/nrc3034. View

20.

Brivanlou A, Gage F, Jaenisch R, Jessell T, Melton D, Rossant J . Stem cells. Setting standards for human embryonic stem cells. Science. 2003; 300(5621):913-6. DOI: 10.1126/science.1082940. View