Cell-Type-Specific Adhesiveness and Proliferation Propensity on Laminin Isoforms Enable Purification of iPSC-Derived Corneal Epithelium

Summary A treatment for intractable diseases is expected to be the replacement of damaged tissues with products from human induced pluripotent stem cells (hiPSCs). Target cell purification is a critical step for realizing hiPSC-based therapy. Here, we found that hiPSC-derived ocular cell types exhibited unique adhesion specificities and growth characteristics on distinct E8 fragments of laminin isoforms (LNE8s): hiPSC-derived corneal epithelial cells (iCECs) and other non-CECs rapidly adhered preferentially to LN332/411/511E8 and LN211E8, respectively, through differential expression of laminin-binding integrins. Furthermore, LN332E8 promoted epithelial cell proliferation but not that of the other eye-related cells, leading to non-CEC elimination by cell competition. Combining these features with magnetic sorting, highly pure iCEC sheets were fabricated. Thus, we established a simple method for isolating iCECs from various hiPSC-derived cells without using fluorescence-activated cell sorting. This study will facilitate efficient manufacture of iCEC sheets for corneal disease treatment and provide insights into target cell-specific scaffold selection.
Highlights • hiPSC-derived cells exhibited differences in expression of laminin-binding integrins • hiPSC-derived corneal epithelial cells (iCECs) adhered to and grew on LN332E8 • Non-epithelial eye-related cells derived from hiPSCs adhered to LN211E8 • Laminin adhesion coupled with magnetic sorting enabled purification of iCEC sheets
Transplantation of hiPSC-derived corneal epithelial cell (iCEC) sheets is expected to serve as a treatment for severe corneal diseases. The efficacy of this treatment depends on effective iCEC purification. Hayashi and colleagues used differential laminin adhesion of iCECs and non-CECs in combination magnetic cell sorting to develop a simple method to obtain highly pure iCEC sheets without using FACS.