Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells
The transplantation of spinal cord progenitor cells (SCPCs) derived from human-induced pluripotent stem cells (iPSCs) has beneficial effects in treating spinal cord injury (SCI). However, the presence of residual undifferentiated iPSCs among their differentiated progeny poses a high risk as these ce...
| Main Authors: | , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2024
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/179447 |
| _version_ | 1826128110730346496 |
|---|---|
| author | Nguyen, Tan Dai Chooi, Wai Hon Jeon, Hyungkook Chen, Jiahui Tan, Jerome Roxby, Daniel N. Lee, Cheryl Yi-Pin Ng, Shi-Yan Chew, Sing Yian Han, Jongyoon |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Nguyen, Tan Dai Chooi, Wai Hon Jeon, Hyungkook Chen, Jiahui Tan, Jerome Roxby, Daniel N. Lee, Cheryl Yi-Pin Ng, Shi-Yan Chew, Sing Yian Han, Jongyoon |
| author_sort | Nguyen, Tan Dai |
| collection | NTU |
| description | The transplantation of spinal cord progenitor cells (SCPCs) derived from human-induced pluripotent stem cells (iPSCs) has beneficial effects in treating spinal cord injury (SCI). However, the presence of residual undifferentiated iPSCs among their differentiated progeny poses a high risk as these cells can develop teratomas or other types of tumors post-transplantation. Despite the need to remove these residual undifferentiated iPSCs, no specific surface markers can identify them for subsequent removal. By profiling the size of SCPCs after a 10-day differentiation process, we found that the large-sized group contains significantly more cells expressing pluripotent markers. In this study, we used a sized-based, label-free separation using an inertial microfluidic-based device to remove tumor-risk cells. The device can reduce the number of undifferentiated cells from an SCPC population with high throughput (ie, >3 million cells/minute) without affecting cell viability and functions. The sorted cells were verified with immunofluorescence staining, flow cytometry analysis, and colony culture assay. We demonstrated the capabilities of our technology to reduce the percentage of OCT4-positive cells. Our technology has great potential for the "downstream processing" of cell manufacturing workflow, ensuring better quality and safety of transplanted cells. |
| first_indexed | 2024-10-01T07:19:34Z |
| format | Journal Article |
| id | ntu-10356/179447 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:19:34Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | ntu-10356/1794472024-08-04T15:38:15Z Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells Nguyen, Tan Dai Chooi, Wai Hon Jeon, Hyungkook Chen, Jiahui Tan, Jerome Roxby, Daniel N. Lee, Cheryl Yi-Pin Ng, Shi-Yan Chew, Sing Yian Han, Jongyoon Lee Kong Chian School of Medicine (LKCMedicine) School of Chemistry, Chemical Engineering and Biotechnology School of Materials Science and Engineering Singapore-MIT Alliance for Research and Technology Centre NTU Institute for Health Technologies Medicine, Health and Life Sciences Cellular therapy Induced pluripotent stem cells The transplantation of spinal cord progenitor cells (SCPCs) derived from human-induced pluripotent stem cells (iPSCs) has beneficial effects in treating spinal cord injury (SCI). However, the presence of residual undifferentiated iPSCs among their differentiated progeny poses a high risk as these cells can develop teratomas or other types of tumors post-transplantation. Despite the need to remove these residual undifferentiated iPSCs, no specific surface markers can identify them for subsequent removal. By profiling the size of SCPCs after a 10-day differentiation process, we found that the large-sized group contains significantly more cells expressing pluripotent markers. In this study, we used a sized-based, label-free separation using an inertial microfluidic-based device to remove tumor-risk cells. The device can reduce the number of undifferentiated cells from an SCPC population with high throughput (ie, >3 million cells/minute) without affecting cell viability and functions. The sorted cells were verified with immunofluorescence staining, flow cytometry analysis, and colony culture assay. We demonstrated the capabilities of our technology to reduce the percentage of OCT4-positive cells. Our technology has great potential for the "downstream processing" of cell manufacturing workflow, ensuring better quality and safety of transplanted cells. National Research Foundation (NRF) Published version This research was supported by the National Research Foundation, Prime Minister’s Office, Singapore under its Campus for Research Excellence and Technological Enterprise (CREATE) program (IntraCREATE grant award number: NRF2019-THE002-0001) and Singapore MIT Alliance for Research and Technology (SMART): Critical Analytics for Manufacturing Personalised Medicine (CAMP) Inter-Disciplinary Research Group. 2024-07-31T06:33:33Z 2024-07-31T06:33:33Z 2024 Journal Article Nguyen, T. D., Chooi, W. H., Jeon, H., Chen, J., Tan, J., Roxby, D. N., Lee, C. Y., Ng, S., Chew, S. Y. & Han, J. (2024). Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells. Stem Cells Translational Medicine, 13(4), 387-398. https://dx.doi.org/10.1093/stcltm/szae002 2157-6564 https://hdl.handle.net/10356/179447 10.1093/stcltm/szae002 38321361 2-s2.0-85190753935 4 13 387 398 en NRF2019-THE002-0001 Stem Cells Translational Medicine © 2024 The Author(s). Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com. application/pdf |
| spellingShingle | Medicine, Health and Life Sciences Cellular therapy Induced pluripotent stem cells Nguyen, Tan Dai Chooi, Wai Hon Jeon, Hyungkook Chen, Jiahui Tan, Jerome Roxby, Daniel N. Lee, Cheryl Yi-Pin Ng, Shi-Yan Chew, Sing Yian Han, Jongyoon Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells |
| title | Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells |
| title_full | Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells |
| title_fullStr | Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells |
| title_full_unstemmed | Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells |
| title_short | Label-free and high-throughput removal of residual undifferentiated cells from iPSC-derived spinal cord progenitor cells |
| title_sort | label free and high throughput removal of residual undifferentiated cells from ipsc derived spinal cord progenitor cells |
| topic | Medicine, Health and Life Sciences Cellular therapy Induced pluripotent stem cells |
| url | https://hdl.handle.net/10356/179447 |
| work_keys_str_mv | AT nguyentandai labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT chooiwaihon labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT jeonhyungkook labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT chenjiahui labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT tanjerome labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT roxbydanieln labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT leecherylyipin labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT ngshiyan labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT chewsingyian labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells AT hanjongyoon labelfreeandhighthroughputremovalofresidualundifferentiatedcellsfromipscderivedspinalcordprogenitorcells |