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  2. Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration

Transplantation of human induced pluripotent stem cell-derived neural crest cells for corneal endothelial regeneration

  • Stem Cell Res Ther. 2021 Mar 29;12(1):214. doi: 10.1186/s13287-021-02267-z.
Yajie Gong 1 2 Haoyun Duan 1 2 Xin Wang 2 3 Can Zhao 1 3 Wenjing Li 2 Chunxiao Dong 2 3 Zongyi Li 4 5 Qingjun Zhou 6 7
Affiliations

Affiliations

  • 1 Shandong First Medical University & Shandong Academy of Medical Sciences, 6699 Qingdao Road, Jinan, 271016, China.
  • 2 State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, 5 Yan'erdao Road, Qingdao, 266071, China.
  • 3 Eye Hospital of Shandong First Medical University, 372 Jingsi Road, Jinan, 250021, China.
  • 4 Shandong First Medical University & Shandong Academy of Medical Sciences, 6699 Qingdao Road, Jinan, 271016, China. [email protected].
  • 5 State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, 5 Yan'erdao Road, Qingdao, 266071, China. [email protected].
  • 6 Shandong First Medical University & Shandong Academy of Medical Sciences, 6699 Qingdao Road, Jinan, 271016, China. [email protected].
  • 7 State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, 5 Yan'erdao Road, Qingdao, 266071, China. [email protected].
Abstract

Background: The corneal endothelium maintains corneal hydration through the barrier and pump function, while its dysfunction may cause corneal edema and vision reduction. Considering its development from neural crest cells (NCCs), here we investigated the efficacy of the human induced pluripotent stem cell (hiPSC)-derived NCCs for corneal endothelial regeneration in rabbits.

Methods: Directed differentiation of hiPSC-derived NCCs was achieved using the chemically defined medium containing GSK-3 Inhibitor and TGF-β inhibitor. The differentiated cells were characterized by immunofluorescence staining, FACS analysis, and in vitro multi-lineage differentiation capacity. For in vivo functional evaluation, 1.0 × 106 hiPSC-derived NCCs or NIH-3 T3 fibroblasts (as control) combined with 100 μM Y-27632 were intracamerally injected into the anterior chamber of rabbits following removal of corneal endothelium. Rabbit corneal thickness and phenotype changes of the transplanted cells were examined at 7 and 14 days with handy pachymeter, dual-immunofluorescence staining, and quantitative RT-PCR.

Results: The hiPSC-derived NCCs were differentiated homogenously through 7 days of induction and exhibited multi-lineage differentiation capacity into peripheral neurons, mesenchymal stem cells, and corneal keratocytes. After 7 days of intracameral injection in rabbit, the hiPSC-derived NCCs led to a gradual recovery of normal corneal thickness and clarity, when comparing to control rabbit with fibroblasts injection. However, the recovery efficacy after 14 days deteriorated and caused the reappearance of corneal edema. Mechanistically, the transplanted cells exhibited the impaired maturation, cellular senescence, and endothelial-mesenchymal transition (EnMT) after the early stage of the in vivo directional differentiation.

Conclusions: Transplantation of the hiPSC-derived NCCs rapidly restored rabbit corneal thickness and clarity. However, the long-term recovery efficacy was impaired by the improper maturation, senescence, and EnMT of the transplanted cells.

Keywords

Corneal endothelium; Neural crest cells; Transplantation; iPSC.

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