Purpose/Aim of the study: To assess if corneal epithelium can be cultured ex-vivo from corneas eviscerated due to irretrievable trauma, according to a cell culture method which made use of autologous platelet-rich plasma (A-PRP) as culture substrate. To compare corneal epithelium cultured ex vivo from corneas eviscerated following trauma using A-PRP combined with DMEM (Dulbecco's modified Eagles medium), versus DMEM alone. Materials and Methods: This was a laboratory case controlled study of human corneal cells cultured in a mixture of A-PRP and DMEM, versus DMEM alone from 6 eviscerated corneas. A hundred explants were created of which fifty explants were plated on A-PRP-gel construct combined with DMEM and fifty controls were placed in serum free DMEM alone. Donor patients received systemic antibiotics prior to evisceration. Results: Confluent epithelium in mono-layers could be cultured when donor limbal biopsies were placed in a mixture of A-PRP culture medium and DMEM. No growth were observed when corneas were placed in serum-free DMEM medium only (p<0.05). No bacterial infection was observed in cultures. Conclusions: This study demonstrated that autologous platelet rich plasma is a viable and effective alternative to bovine serum for the ex-vivo expansion of limbal epithelial cells. It also shows that eviscerated corneas are a viable source of donor tissue for this purpose in South Africa where access to tissue banks is limited.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:uct/oai:localhost:11427/22923 |
| Date | January 2016 |
| Creators | Heydenrych, Leonard Goussárd |
| Contributors | Du Toit, D F |
| Publisher | University of Cape Town, Faculty of Health Sciences, Division of Ophthalmology |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Master Thesis, Masters, MMed |
| Format | application/pdf |
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