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Biotechnol Lett ; 43(6): 1241-1251, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33768381

RESUMO

PURPOSE: McCarey-Kaufman's (MK) medium and Optisol-GS medium are the most commonly employed media for human donor corneal preservation. In this study, we evaluated the preservation efficacy of discarded human donor corneas using a Thermo-reversible gelation polymer (TGP) added to these two media. METHODS: Thirteen human corneal buttons collected from deceased donors, which were otherwise discarded due to low endothelial cell density (ECD) were used. They were stored in four groups: MK medium, MK medium with TGP, Optisol-GS and Optisol-GS with TGP at 4 °C for 96 h. Slit lamp examination and specular microscopy were performed. Corneal limbal tissues from these corneas were then cultured using explant methodology one with and the other without TGP scaffold, for 21 days. RESULTS: MK + TGP and Optisol-GS + TGP preserved corneas better than without TGP, which was observed by maintenance of ECD which was significantly higher in Optisol-GS + TGP than MK + TGP (p-value = 0.000478) and corneal thickness remaining the same for 96 h. Viable corneal epithelial cells could be grown from the corneas stored only in MK + TGP and Optisol-GS + TGP. During culture, the TGP scaffold helped maintain the native epithelial phenotype and progenitor/stem cell growth was confirmed by RT-PCR characterization. CONCLUSION: TGP reconstituted with MK and Optisol-GS media yields better preservation of human corneal buttons in terms of relatively higher ECD maintenance and better in vitro culture outcome of corneal limbal tissue. This method has the potential to become a standard donor corneal transportation-preservation methodology and it can also be extended to other tissue or organ transportation upon further validation.


Assuntos
Meios de Cultura/química , Endotélio Corneano/citologia , Preservação de Tecido/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Cadáver , Sulfatos de Condroitina/química , Misturas Complexas/química , Dextranos/química , Feminino , Gentamicinas/química , Humanos , Masculino , Pessoa de Meia-Idade , Compostos Orgânicos/química , Microscopia com Lâmpada de Fenda
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