Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation
The culture of human corneal endothelial cells (CECs) is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the prop...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | International Journal of Biomaterials |
| Online Access: | http://dx.doi.org/10.1155/2012/601302 |
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| author | Gary S. L. Peh Man-Xin Lee Fei-Yi Wu Kah-Peng Toh Deepashree Balehosur Jodhbir S. Mehta |
| author_facet | Gary S. L. Peh Man-Xin Lee Fei-Yi Wu Kah-Peng Toh Deepashree Balehosur Jodhbir S. Mehta |
| author_sort | Gary S. L. Peh |
| collection | DOAJ |
| description | The culture of human corneal endothelial cells (CECs) is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the propagation of slow proliferative CECs in vitro can be hindered by rapid growing stromal corneal fibroblasts (CSFs) that may be coisolated in some cases. The purpose of this study was to evaluate a strategy using magnetic cell separation (MACS) technique to deplete the contaminating CSFs from CEC cultures using antifibroblast magnetic microbeads. Separated “labeled” and “flow-through” cell fractions were collected separately, cultured, and morphologically assessed. Cells from the “flow-through” fraction displayed compact polygonal morphology and expressed Na+/K+ATPase indicative of corneal endothelial cells, whilst cells from the “labeled” fraction were mostly elongated and fibroblastic. A separation efficacy of 96.88% was observed. Hence, MACS technique can be useful in the depletion of contaminating CSFs from within a culture of CECs. |
| format | Article |
| id | doaj-art-4f40425e15b0409b8d549c0df511a0ba |
| institution | Kabale University |
| issn | 1687-8787 1687-8795 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Biomaterials |
| spelling | doaj-art-4f40425e15b0409b8d549c0df511a0ba2025-02-03T05:47:35ZengWileyInternational Journal of Biomaterials1687-87871687-87952012-01-01201210.1155/2012/601302601302Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell SeparationGary S. L. Peh0Man-Xin Lee1Fei-Yi Wu2Kah-Peng Toh3Deepashree Balehosur4Jodhbir S. Mehta5Singapore Eye Research Institute, 168751, SingaporeSingapore Eye Research Institute, 168751, SingaporeSingapore Eye Research Institute, 168751, SingaporeSingapore Eye Research Institute, 168751, SingaporeSingapore Eye Research Institute, 168751, SingaporeSingapore Eye Research Institute, 168751, SingaporeThe culture of human corneal endothelial cells (CECs) is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the propagation of slow proliferative CECs in vitro can be hindered by rapid growing stromal corneal fibroblasts (CSFs) that may be coisolated in some cases. The purpose of this study was to evaluate a strategy using magnetic cell separation (MACS) technique to deplete the contaminating CSFs from CEC cultures using antifibroblast magnetic microbeads. Separated “labeled” and “flow-through” cell fractions were collected separately, cultured, and morphologically assessed. Cells from the “flow-through” fraction displayed compact polygonal morphology and expressed Na+/K+ATPase indicative of corneal endothelial cells, whilst cells from the “labeled” fraction were mostly elongated and fibroblastic. A separation efficacy of 96.88% was observed. Hence, MACS technique can be useful in the depletion of contaminating CSFs from within a culture of CECs.http://dx.doi.org/10.1155/2012/601302 |
| spellingShingle | Gary S. L. Peh Man-Xin Lee Fei-Yi Wu Kah-Peng Toh Deepashree Balehosur Jodhbir S. Mehta Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation International Journal of Biomaterials |
| title | Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation |
| title_full | Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation |
| title_fullStr | Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation |
| title_full_unstemmed | Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation |
| title_short | Optimization of Human Corneal Endothelial Cells for Culture: The Removal of Corneal Stromal Fibroblast Contamination Using Magnetic Cell Separation |
| title_sort | optimization of human corneal endothelial cells for culture the removal of corneal stromal fibroblast contamination using magnetic cell separation |
| url | http://dx.doi.org/10.1155/2012/601302 |
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