A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin
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...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-06-01
|
| Series: | Regenerative Therapy |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352320420300419 |
| _version_ | 1818039042595356672 |
|---|---|
| author | Yoshikazu Haramoto Yasuko Onuma Shuuji Mawaribuchi Yoshiro Nakajima Yasuhiko Aiki Kumiko Higuchi Madoka Shimizu Hiroaki Tateno Jun Hirabayashi Yuzuru Ito |
| author_facet | Yoshikazu Haramoto Yasuko Onuma Shuuji Mawaribuchi Yoshiro Nakajima Yasuhiko Aiki Kumiko Higuchi Madoka Shimizu Hiroaki Tateno Jun Hirabayashi Yuzuru Ito |
| author_sort | Yoshikazu Haramoto |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-10T07:52:21Z |
| format | Article |
| id | doaj.art-f96df89f1fe7470286eb8cce3bbb1c39 |
| institution | Directory Open Access Journal |
| issn | 2352-3204 |
| language | English |
| last_indexed | 2024-12-10T07:52:21Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj.art-f96df89f1fe7470286eb8cce3bbb1c392022-12-22T01:57:00ZengElsevierRegenerative Therapy2352-32042020-06-0114306314A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectinYoshikazu Haramoto0Yasuko Onuma1Shuuji Mawaribuchi2Yoshiro Nakajima3Yasuhiko Aiki4Kumiko Higuchi5Madoka Shimizu6Hiroaki Tateno7Jun Hirabayashi8Yuzuru Ito9Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, JapanDivision of Developmental Biology, Department of Anatomy, Kyoto Prefectural University of Medicine, Kawaramachi, Hirokoji, Kamigyo-ku, Kyoto, 602-8566, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 2, 1-1-1, Umezono, Tsukuba, Ibaraki, 305-8568, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 2, 1-1-1, Umezono, Tsukuba, Ibaraki, 305-8568, JapanBiotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan; Corresponding author.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.http://www.sciencedirect.com/science/article/pii/S2352320420300419Pluripotent stem cellrBC2LCNLectinCell separation |
| spellingShingle | Yoshikazu Haramoto Yasuko Onuma Shuuji Mawaribuchi Yoshiro Nakajima Yasuhiko Aiki Kumiko Higuchi Madoka Shimizu Hiroaki Tateno Jun Hirabayashi Yuzuru Ito A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin Regenerative Therapy Pluripotent stem cell rBC2LCN Lectin Cell separation |
| title | A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin |
| title_full | A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin |
| title_fullStr | A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin |
| title_full_unstemmed | A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin |
| title_short | A technique for removing tumourigenic pluripotent stem cells using rBC2LCN lectin |
| title_sort | technique for removing tumourigenic pluripotent stem cells using rbc2lcn lectin |
| topic | Pluripotent stem cell rBC2LCN Lectin Cell separation |
| url | http://www.sciencedirect.com/science/article/pii/S2352320420300419 |
| work_keys_str_mv | AT yoshikazuharamoto atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yasukoonuma atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT shuujimawaribuchi atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yoshironakajima atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yasuhikoaiki atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT kumikohiguchi atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT madokashimizu atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT hiroakitateno atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT junhirabayashi atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yuzuruito atechniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yoshikazuharamoto techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yasukoonuma techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT shuujimawaribuchi techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yoshironakajima techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yasuhikoaiki techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT kumikohiguchi techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT madokashimizu techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT hiroakitateno techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT junhirabayashi techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin AT yuzuruito techniqueforremovingtumourigenicpluripotentstemcellsusingrbc2lcnlectin |