Optimized human platelet lysate as novel basis for a serum-, xeno- and additive-free corneal endothelial cell and tissue culture

Thieme D, Reuland L, Lindl T, Kruse F, Fuchsluger T (2017)

Publication Type: Journal article

Publication year: 2017

Journal

Journal of Tissue Engineering and Regenerative Medicine Wiley-Blackwell

DOI: 10.1002/term.2574

Abstract

The expansion of donor derived corneal endothelial cells is a promising approach for regenerative therapies in corneal diseases. To achieve the best GMP standard the entire cultivation process should be devoid of non-human components. However, so far there is no suitable xeno-free protocol for clinical applications.We therefore introduce a processed variant of a platelet lysate for the use in corneal cell and tissue culture based on a GMP-grade thrombocyte concentrate. This processed human platelet lysate (phPL), free of any animal components and of anti-coagulants like heparin with a physiological ionic composition, was used to cultivate corneal endothelial cells (EC) in vitro and ex vivo in comparison to standard cultivation with FCS. Human donor corneas were cut in quarters while two quarters of each cornea were incubated with the respective medium supplement. Three fields of view per quarter were taken into account for the analysis.Evaluation of phPL as a medium supplement in cell culture of immortalized EC showed a superior viability compared to fetal calf serum (FCS) control with reduced cell proliferation. Furthermore, the viability during the expansion of primary cells is significantly (3fold+-0.5) increased with phPL compared to FCS standard medium. Quartering donor corneas was traumatic for the endothelium and therefore resulted in increased EC loss. Interestingly, however, cultivation of the quartered pieces for two weeks in 0.1mg/mL pHPL in Biochrome I showed a 21 (+-10) % EC loss compared to 67 (+-12) % EC loss when cultivated in 2% FCS in Biochrome I.The cell culture protocol with pHPL as FCS replacement seems to be superior to the standard FCS protocols with respect to EC survival. It offers a xeno-free and physiological environment for corneal endothelial cells. This alternative cultivation protocol could facilitate the use of EC for human corneal cell therapy.

Authors with CRIS profile

Friedrich Kruse Lehrstuhl für Augenheilkunde Thomas Fuchsluger Professur für Augenheilkunde (Cornea/vorderer Augenabschnitt)

Involved external institutions

IAZ-GmbH DE Germany (DE)

How to cite

APA:

Thieme, D., Reuland, L., Lindl, T., Kruse, F., & Fuchsluger, T. (2017). Optimized human platelet lysate as novel basis for a serum-, xeno- and additive-free corneal endothelial cell and tissue culture. Journal of Tissue Engineering and Regenerative Medicine. https://dx.doi.org/10.1002/term.2574

MLA:

Thieme, Daniel, et al. "Optimized human platelet lysate as novel basis for a serum-, xeno- and additive-free corneal endothelial cell and tissue culture." Journal of Tissue Engineering and Regenerative Medicine (2017).

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