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

• Published in: Stem cell reports Vol. 14; no. 4; pp. 663 - 676
• Main Authors: Shibata, Shun, Hayashi, Ryuhei, Kudo, Yuji, Okubo, Toru, Imaizumi, Tsutomu, Katayama, Tomohiko, Ishikawa, Yuki, Kobayashi, Yuki, Toga, Junko, Taniguchi, Yukimasa, Honma, Yoichi, Sekiguchi, Kiyotoshi, Nishida, Kohji
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 14.04.2020; Elsevier
• Subjects: Cell Adhesion - drug effects; cell competition; Cell Differentiation - drug effects; Cell Proliferation - drug effects; cell purification; Cell Separation - methods; Cells, Cultured; corneal epithelial cells (CECs); Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Epithelium, Corneal - cytology; human induced pluripotent stem cells (hiPSCs); Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - drug effects; Integrins - metabolism; Laminin - pharmacology; laminin isoforms; Protein Isoforms - pharmacology; substrate-specific adhesion; substrate-specific adhesion; cell purification; cell competition; laminin isoforms; human induced pluripotent stem cells (hiPSCs); corneal epithelial cells (CECs)
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2020.02.008

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. •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.