Abstract
PurposeSeeking to improve the access to regenerative medicine, this study investigated the structural and transcriptional effects of storage temperature on human oral mucosal epithelial cells (OMECs).MethodsCells were stored at four different temperatures (4°C, 12°C, 24°C and 37°C) for two weeks. Then, the morphology, cell viability and differential gene expression were examined using light and scanning electron microscopy, trypan blue exclusion test and TaqMan gene expression array cards, respectively.ResultsCells stored at 4°C had the most similar morphology to non-stored controls with the highest viability rate (58%), whereas the 37°C group was most dissimilar with no living cells. The genes involved in stress-induced growth arrest (GADD45B) and cell proliferation inhibition (TGFB2) were upregulated at 12°C and 24°C. Upregulation was also observed in multifunctional genes responsible for morphology, growth, adhesion and motility such as EFEMP1 (12°C) and EPHA4 (4°C–24°C). Among genes used as differentiation markers, PPARA and TP53 (along with its associated gene CDKN1A) were downregulated in all temperature conditions, whereas KRT1 and KRT10 were either unchanged (4°C) or downregulated (24°C and 12°C; and 24°C, respectively), except for upregulation at 12°C for KRT1.ConclusionsCells stored at 12°C and 24°C were stressed, although the expression levels of some adhesion-, growth- and apoptosis-related genes were favourable. Collectively, this study suggests that 4°C is the optimal storage temperature for maintenance of structure, viability and function of OMECs after two weeks.
Highlights
Limbal stem cell deficiency (LSCD) is a potentially painful and blinding condition caused by damage or loss of the limbal stem cells [1, 2]
Cells stored at 4 ̊C had the most similar morphology to non-stored controls with the highest viability rate (58%), whereas the 37 ̊C group was most dissimilar with no living cells
This study suggests that 4 ̊C is the optimal storage temperature for maintenance of structure, viability and function of Oral mucosal epithelial cells (OMECs) after two weeks
Summary
Limbal stem cell deficiency (LSCD) is a potentially painful and blinding condition caused by damage or loss of the limbal stem cells [1, 2]. Cell therapy has gained increasing attention over the last few years in the treatment of LSCD after the first successful transplantation of autologous limbal stem cells in 1997 by Italian scientists for patients with unilateral disease [3]. This therapy is believed to replace or restore remaining functioning limbal stem cells to promote regeneration of the corneal epithelium [4]. Oral mucosal epithelial cells (OMECs) were the first alternative autologous source to be studied in both rabbits [5] and humans [6]. Seeking to improve transplantation success and expand access to regenerative medicine, researchers have investigated different storage conditions [10,11,12,13,14,15,16] and transportation techniques [17,18,19]
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