Markers for Distinguishing Cultured Human Corneal Endothelial Cells from Corneal Stromal Myofibroblasts

作者：Yamaguchi Masahiro; Shima Nobuyuki^*; Kimoto Miwa; Ebihara Nobuyuki; Murakami Akira; Yamagami Satoru

来源：Current Eye Research, 2015, 40(12): 1211-1217.

DOI：10.3109/02713683.2014.993087

• Corneal endothelial cell
• differential marker
• keratocyte
• myofibroblast
• regenerative medicine

Purpose: Eliminating contamination by corneal stromal cells is critical when preparing cultured human corneal endothelial cells (CECs) transplantation. We investigated markers for the purification of cultured human CECs and markers for excluding cultured human corneal stromal myofibroblasts (CSMFs) from cultured human CECs.
Materials and methods: CECs and CSMFs were obtained from human donor corneas by culturing separately in serum-containing medium. Candidate markers of CECs and CSMFs were screened with microarray analysis in the fourth passaged CECs and CSMFs. Then, selected factors were evaluated in reverse transcription polymerase chain reaction (RT-PCR), western blot, immunocytochemistry, and flow cytometry to investigate differential markers for each cell.
Results: Among the genes identified by microarray analysis, cultured human CECs, but not CSMFs, expressed integrin alpha 3 (ITGA3 and CD49c) protein according to immunocytochemistry and western blotting. Iroquois homeobox 2 (IRX2) gene was a marker that distinguished CSMFs from cultured human CECs by RT-PCR. The IRX2 gene can be used as a marker of CSMFs contaminating cultured CECs.
Conclusion: These molecules could be important markers for the production of highly purified cultured CECs for regenerative medicine.

• 出版日期2015