The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach
The global shortage of donor corneas has garnered extensive interest in the development of graft alternatives suitable for endothelial keratoplasty using cultivated primary human corneal endothelial cells (CECs). We have recently described a dual media approach for the propagation of human CECs. In...
| Main Authors: | , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2015
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/107251 http://hdl.handle.net/10220/25528 |
| _version_ | 1826129177142624256 |
|---|---|
| author | Peh, Gary S. L. Adnan, Khadijah George, Benjamin L. Ang, Heng Pei Seah, Xin Yi Tan, Donald T. Mehta, Jodhbir Singh |
| author2 | School of Materials Science & Engineering |
| author_facet | School of Materials Science & Engineering Peh, Gary S. L. Adnan, Khadijah George, Benjamin L. Ang, Heng Pei Seah, Xin Yi Tan, Donald T. Mehta, Jodhbir Singh |
| author_sort | Peh, Gary S. L. |
| collection | NTU |
| description | The global shortage of donor corneas has garnered extensive interest in the development of graft alternatives suitable for endothelial keratoplasty using cultivated primary human corneal endothelial cells (CECs). We have recently described a dual media approach for the propagation of human CECs. In this work, we characterize the effects of a Rho-kinase inhibitor Y-27632 on the cultivation of CECs propagated using the dual media culture system. Seventy donor corneas deemed unsuitable for transplantation were procured for this study. We assessed the use of Y-27632 for its effect at each stage of the cell culture process, specifically for cell attachment, cell proliferation, and during both regular passaging and cryopreservation. Lastly, comparison of donor-matched CEC-cultures expanded with or without Y-27632 was also performed. Our results showed that Y-27632 significantly improved the attachment and proliferation of primary CECs. A non-significant pro-survival effect was detected during regular cellular passage when CECs were pre-treated with Y-27632, an effect that became more evident during cryopreservation. Our study showed that the inclusion of Y-27632 was beneficial for the propagation of primary CECs expanded via the dual media approach, and was able to increase overall cell yield by between 1.96 to 3.36 fold. |
| first_indexed | 2024-10-01T07:36:26Z |
| format | Journal Article |
| id | ntu-10356/107251 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:36:26Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | ntu-10356/1072512023-07-14T15:45:44Z The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach Peh, Gary S. L. Adnan, Khadijah George, Benjamin L. Ang, Heng Pei Seah, Xin Yi Tan, Donald T. Mehta, Jodhbir Singh School of Materials Science & Engineering DRNTU::Science::Biological sciences::Human anatomy and physiology The global shortage of donor corneas has garnered extensive interest in the development of graft alternatives suitable for endothelial keratoplasty using cultivated primary human corneal endothelial cells (CECs). We have recently described a dual media approach for the propagation of human CECs. In this work, we characterize the effects of a Rho-kinase inhibitor Y-27632 on the cultivation of CECs propagated using the dual media culture system. Seventy donor corneas deemed unsuitable for transplantation were procured for this study. We assessed the use of Y-27632 for its effect at each stage of the cell culture process, specifically for cell attachment, cell proliferation, and during both regular passaging and cryopreservation. Lastly, comparison of donor-matched CEC-cultures expanded with or without Y-27632 was also performed. Our results showed that Y-27632 significantly improved the attachment and proliferation of primary CECs. A non-significant pro-survival effect was detected during regular cellular passage when CECs were pre-treated with Y-27632, an effect that became more evident during cryopreservation. Our study showed that the inclusion of Y-27632 was beneficial for the propagation of primary CECs expanded via the dual media approach, and was able to increase overall cell yield by between 1.96 to 3.36 fold. Published version 2015-05-14T03:03:46Z 2019-12-06T22:27:23Z 2015-05-14T03:03:46Z 2019-12-06T22:27:23Z 2015 2015 Journal Article Peh, G. S. L., Adnan, K., George, B. L., Ang, H. P., Seah, X. Y., Tan, D. T., et al. (2015). The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach. Scientific Reports, 5, 9167-. 2045-2322 https://hdl.handle.net/10356/107251 http://hdl.handle.net/10220/25528 10.1038/srep09167 25823914 en Scientific reports This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Human anatomy and physiology Peh, Gary S. L. Adnan, Khadijah George, Benjamin L. Ang, Heng Pei Seah, Xin Yi Tan, Donald T. Mehta, Jodhbir Singh The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach |
| title | The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach |
| title_full | The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach |
| title_fullStr | The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach |
| title_full_unstemmed | The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach |
| title_short | The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach |
| title_sort | effects of rho associated kinase inhibitor y 27632 on primary human corneal endothelial cells propagated using a dual media approach |
| topic | DRNTU::Science::Biological sciences::Human anatomy and physiology |
| url | https://hdl.handle.net/10356/107251 http://hdl.handle.net/10220/25528 |
| work_keys_str_mv | AT pehgarysl theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT adnankhadijah theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT georgebenjaminl theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT anghengpei theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT seahxinyi theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT tandonaldt theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT mehtajodhbirsingh theeffectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT pehgarysl effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT adnankhadijah effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT georgebenjaminl effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT anghengpei effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT seahxinyi effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT tandonaldt effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach AT mehtajodhbirsingh effectsofrhoassociatedkinaseinhibitory27632onprimaryhumancornealendothelialcellspropagatedusingadualmediaapproach |