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Identification and characterization of dendritic cell subtypes in preserved and cultured cadaveric human corneolimbal tissue on amniotic membrane
Author(s) -
Lužnik Zala,
Kopitar Andreja Nataša,
Lapajne Luka,
Pižem Jože,
Ferrari Stefano,
Ihan Alojz,
Hawlina Marko,
Schollmayer Petra
Publication year - 2019
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/aos.13854
Subject(s) - explant culture , biology , cd86 , cd80 , stromal cell , stem cell , ex vivo , cd11c , cell sorting , stem cell marker , flow cytometry , immunology , progenitor cell , microbiology and biotechnology , andrology , pathology , t cell , in vivo , in vitro , immune system , medicine , cd40 , cancer research , phenotype , biochemistry , cytotoxic t cell , gene
Purpose Rejection is the leading cause of failure of limbal allogafts. Resident dendritic cell ( DC ) maturation plays a critical role in host allosensitization. There are two lineages: myeloid ( mDC ) and lymphoid ( pDC ), with different biological properties. The aim was to analyse the distribution of DC subtypes in limbal explant cultures on amniotic membrane ( AM ), cultivated on either the epithelial or stromal side and to compare the results with directly isolated cells from cadaveric whole corneoscleral tissue divided into specific areas. Methods The expression of CD 11c ( mDC ), CD 303/ CD 123 ( pDC ) and costimulatory molecules CD 80, CD 86 and activation markers HLA ‐ DR , CD 83 was investigated by flow cytometry. Additionally, the corneal epithelium marker CK 12 and ABCB 5, a new epithelial stem cell marker, were investigated. Results Cells positive for pDC and mDC markers were found in all examined areas, with a nonsignificant prevalence of pDC . In limbal explant cultures on AM , the percentage of pDC and mDC was similar, with no statistically significant difference between cultures on epithelial or stromal sides of AM . However, with ex vivo limbal explant cultivation on AM , the pDC content declined significantly (p < 0.05) and the ABCB 5 marker was likewise statistically significantly reduced. Conclusion This is the first study to characterize the distribution of pDC and mDC subsets in cultured and noncultured human corneolimbal tissue. Additionally, ABCB 5 positive cells were identified. These findings might be important for future strategies, allowing preparation of corneolimbal allografts with optimal stem cell content for a longer lasting therapeutic effect.