Eviscerated corneas as tissue source for ex vivo expansion of limbal epithelial cells on platelet-rich plasma gels

Master Thesis


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University of Cape Town

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.