Cryoprotectants for the vitrification of corneal endothelial cells.
The objective of this work was to select and test systematically possible cryoprotective agents (CPAs) and to obtain a suitable formula for vitrification of corneal endothelial cells (CECs). Fresh bovine CECs were isolated and tested with an optimized vitrification protocol with multi-step CPA loadi...
| Main Authors: | , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2009
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| _version_ | 1797051741184720896 |
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| author | Fan, W Ma, X Ge, D Liu, T Cui, Z |
| author_facet | Fan, W Ma, X Ge, D Liu, T Cui, Z |
| author_sort | Fan, W |
| collection | OXFORD |
| description | The objective of this work was to select and test systematically possible cryoprotective agents (CPAs) and to obtain a suitable formula for vitrification of corneal endothelial cells (CECs). Fresh bovine CECs were isolated and tested with an optimized vitrification protocol with multi-step CPA loading and removal. Three types of CPAs components, i.e. the penetrating CPAs, sugars and macromolecular compounds, were experimentally evaluated using the viability assayed by trypan blue. Dimethyl sulfoxide, ethylene glycol (EG), 1,2-propanediol, 2,3-butanediol, acetamide and ethylene glycol monomethyl ether were chosen as the penetrating CPA components. Sugars including xylose, fructose, mannose, glucose, maltose, sucrose and trehalose were tested. Ficoll (MW 7kDa), dextran (MW 7kDa), chondroitin sulfate (CS, MW 18-30kDa), bovine serum albumin (MW 68kDa) and polyethylene glycol (MW 6kDa, 10kDa and 20kDa) were chosen as the macromolecular compounds. CECs were also preserved by slow freezing as a control. The results showed that EG was the most suitable penetrating CPA component and glucose the most suitable sugar, and CS the most suitable macromolecule. The optimized concentrations for each component in the vitrification solution were 52% (w/w) EG, 8% (w/w) glucose and 3% (w/w) CS. The CEC survival rate of 89.4+/-2.1% (mean+/-SD) was obtained using this formula and established vitrification protocol which was comparable to that by slow freezing. |
| first_indexed | 2024-03-06T18:23:48Z |
| format | Journal article |
| id | oxford-uuid:073abbcc-6bcb-4628-b5f7-eb37907fe0b4 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T18:23:48Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | oxford-uuid:073abbcc-6bcb-4628-b5f7-eb37907fe0b42022-03-26T09:06:29ZCryoprotectants for the vitrification of corneal endothelial cells.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:073abbcc-6bcb-4628-b5f7-eb37907fe0b4EnglishSymplectic Elements at Oxford2009Fan, WMa, XGe, DLiu, TCui, ZThe objective of this work was to select and test systematically possible cryoprotective agents (CPAs) and to obtain a suitable formula for vitrification of corneal endothelial cells (CECs). Fresh bovine CECs were isolated and tested with an optimized vitrification protocol with multi-step CPA loading and removal. Three types of CPAs components, i.e. the penetrating CPAs, sugars and macromolecular compounds, were experimentally evaluated using the viability assayed by trypan blue. Dimethyl sulfoxide, ethylene glycol (EG), 1,2-propanediol, 2,3-butanediol, acetamide and ethylene glycol monomethyl ether were chosen as the penetrating CPA components. Sugars including xylose, fructose, mannose, glucose, maltose, sucrose and trehalose were tested. Ficoll (MW 7kDa), dextran (MW 7kDa), chondroitin sulfate (CS, MW 18-30kDa), bovine serum albumin (MW 68kDa) and polyethylene glycol (MW 6kDa, 10kDa and 20kDa) were chosen as the macromolecular compounds. CECs were also preserved by slow freezing as a control. The results showed that EG was the most suitable penetrating CPA component and glucose the most suitable sugar, and CS the most suitable macromolecule. The optimized concentrations for each component in the vitrification solution were 52% (w/w) EG, 8% (w/w) glucose and 3% (w/w) CS. The CEC survival rate of 89.4+/-2.1% (mean+/-SD) was obtained using this formula and established vitrification protocol which was comparable to that by slow freezing. |
| spellingShingle | Fan, W Ma, X Ge, D Liu, T Cui, Z Cryoprotectants for the vitrification of corneal endothelial cells. |
| title | Cryoprotectants for the vitrification of corneal endothelial cells. |
| title_full | Cryoprotectants for the vitrification of corneal endothelial cells. |
| title_fullStr | Cryoprotectants for the vitrification of corneal endothelial cells. |
| title_full_unstemmed | Cryoprotectants for the vitrification of corneal endothelial cells. |
| title_short | Cryoprotectants for the vitrification of corneal endothelial cells. |
| title_sort | cryoprotectants for the vitrification of corneal endothelial cells |
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