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Steffi Matthyssen, Veerle Van Gerwen, Carina Koppen, Nadia Zakaria; Comparative genetic analysis of human mesenchymal stem cells isolated from corneal stroma and bone marrow. Invest. Ophthalmol. Vis. Sci. 2018;59(9):2276. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal stroma-derived mesenchymal stem cells (CS-MSC) have not yet been directly compared on a genetic level with bone marrow MSC (BM-MSC), with or without the inclusion of xenofree conditions. This results in inconclusiveness with regards to the MSC-like properties of corneal stromal cells. The potential of corneal-derived MSCs in tissue engineering applications for Ophthalmology is yet to be explored. We hypothesize that CS-MSC show comparable expression of key MSC genes to BM-MSC, in accordance to findings on a phenotypical and functional level, and that xenofree culture does not alter genetic expression patterns in CS-MSC.
Human CS-MSC were obtained from six donors and expanded in both standard conditions (basal medium with the addition of 10% Fetal Bovine Serum – FBS) and xenofree conditions using 10% Human Platelet Lysate (HPL) as a replacement for FBS. At passage 4, the cells were harvested and RNA isolation was followed by cDNA extraction and qPCR for 85 key genes for human MSC. Human BM-MSCs at passage 6 (provided by Veneto Eye Bank) were used as control. Phenotypic characterization using flow cytometry and trilineage differentiation was performed to confirm the MSC-like properties of corneal stroma-derived cells as required by the International Society for Cellular Therapy (ISCT).
Out of 85 genes tested, 71 showed no differential expression between bone marrow-derived and corneal stroma-derived MSCs in normal or xenofree conditions. Of the 14 genes where expression differed between BM- and CS-MSCs, 12 were not expressed in BM. Genes with variable expression include: ABCB1, IL10, PROM1, CSF3, IL1B, TNF, KDR, SOX2, BMP7, HNF1A, FGF10 and WNT3A. Flow cytometry showed that corneal stroma-derived MSCs, like BM-SCs, met the ISCT criteria and were positive for CD73, CD90, CD105, CD13, CD29, CD44, CD166 and negative for CD11b, CD 14, CD19, CD34, CD45, CD79a, HLA-DR and were able to differentiate into osteocytes, chondrocytes and adipocytes regardless of the cultures being xenofree or not.
Despite portraying an identical phenotype as described by the ISCT criteria, MSCs derived from the corneal stroma show a different genetic profile to those that are derived from bone marrow. Further research is warranted in order to determine if CS-MSCs would be superior to BM-MSCs for corneal stromal tissue engineering.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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