Abstract
Purpose To ex vivo expand oral mucosal epithelium cells (OMECs) on acellular porcine corneal stroma (APCS) without using feeder cells and serum and to compare the morphologic and phenotypic characteristics of cultured oral cells on APCS to those of cells on deluded human amniotic membrane (HAM). Methods SD rat oral mucosal biopsies were cultured on APCS and HAM. Reverse-transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to analyze the characterization of stem cells and epithelial differentiation of the outgrowth products. Results Stratified and optimal transplantable OMECs were obtained after being cultured three to four weeks. Both RT-PCR and immunohistochemistry showed that cultured OMECs expressed markers of epithelial differentiation cytokeratin K3 and epithelial stem cell markers of p63 and ABCG2. Conclusions OMECs can be successfully cultured on APCS without using xenobiotic feeder cells and serum. Characterization showed that these sheets retain the morphologic and phenotypic characteristics of OMECs within differentiated cells and stem cells. The optimal transplantable sheets can prove to be particularly beneficial to both bilateral limbal stem cell deficiency and deep corneal lesions.
Highlights
Limbal stem cells (LSCs) have been proven with two main functions: renewed corneal epithelial cells and conjunctival and blood vessel ingrowth onto the cornea
Located at the basal layer of the limbal epithelium [1, 2], it could be fully destroyed by severe ocular surface diseases (OSD), including chemical burns, microbiological infections, autoimmune diseases, and genetic disorders, which results in limbal stem cell deficiency (LSCD)
To investigate the feasibility of Acellular porcine corneal stroma (APCS) as a carrier for ex vivo oral mucosal epithelium cells (OMECs) growth, we observed the growth of cells and designed Reverse-transcription polymerase chain reaction (RT-polymerase chain reaction (PCR)) and immunocytochemistry to explore the morphology and phenotype of COMECs
Summary
Limbal stem cells (LSCs) have been proven with two main functions: renewed corneal epithelial cells and conjunctival and blood vessel ingrowth onto the cornea. COMECs were mainly used in superficial corneal diseases, such as corneas covered by a thin vascular membrane, with or without the substrate and carrier such as contact lenses (CLs) [26] and human amniotic membrane (HAM) [10, 27,28,29,30,31,32,33,34,35]. If patients have both LSCD and deep corneal cloudiness, keratoplasty should be followed by the COMECs transplantation.
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