Journal: The Journal of Clinical Investigation

Article Title: Long-term corneal recovery by simultaneous delivery of hPSC-derived corneal endothelial precursors and nicotinamide

doi: 10.1172/JCI146658

Figure Lengend Snippet: ( A ) Human pCECs and hESC-derived NCCs, CEPs, and CECs were intracamerally injected in the rabbit corneal endothelial dysfunction model. Corneal transparency was assessed by slit-lamp microscopy after 1, 3, 7, and 14 days of transplantation. ( B ) Central corneal thicknesses were measured by pachymeter after 1, 3, 7, 10, and 14 days of transplantation. n = 3. The dashed line shows normal corneal thickness. ( C ) Immunostaining of ZO-1 and ATP1A1 in the transplanted CEPs 7 days after transplantation. The transplanted cells were stained by human specific antibody human nuclei (HuNu). Nuclei were stained with DAPI. Scale bar: 50 μm. ( D ) Expressions of EnMT markers in the pretransplanted (Pre) and transplanted CEPs 7 days after transplantation by qPCR. n = 3. *P < 0.05, **P < 0.01 by 2-tailed Student’s t test. ( E ) Immunostaining of α-SMA and fibronectin in the transplanted CEPs 7 days after transplantation. The transplanted cells were stained by HuNu. Nuclei were stained with DAPI. Scale bar: 50 μm.

Article Snippet: The cells were viewed under bright field microscopy (Echo Laboratories).

Techniques: Derivative Assay, Injection, Microscopy, Transplantation Assay, Immunostaining, Staining

Journal: The Journal of Clinical Investigation

Article Title: Long-term corneal recovery by simultaneous delivery of hPSC-derived corneal endothelial precursors and nicotinamide

doi: 10.1172/JCI146658

Figure Lengend Snippet: ( A ) Schema of the intracameral injection of hESC-derived CEPs and NAM treatment in the rabbit model of corneal endothelial dysfunction. ( B ) Corneal transparency was assessed by slit-lamp microscopy in rabbits with simple cell injection (CEP), cell injection and NAM treatment (CEP+NAM), and NAM alone treatment (NAM) 1, 3, 7, and 14 days after transplantation. ( C ) Central corneal thicknesses were measured by pachymeter 1, 3, 7, 10, and 14 days after transplantation. n = 6. The dashed line shows normal corneal thickness. ( D ) Regenerated corneal endothelium was evaluated by corneal confocal microscopy 7 days after transplantation, with the normal rabbit corneal endothelium (Norm) as control. ( E ) Statistical analysis of corneal endothelial density 7 days after transplantation. n = 6 **P <0.01 by 1-way ANOVA with Tukey’s HSD test. ( F ) Double staining of F-actin and human cell surface determinant TRA-1-85 and HuNu 14 days after transplantation. Nuclei were stained with DAPI. Scale bar: 50 μm.

Article Snippet: The cells were viewed under bright field microscopy (Echo Laboratories).

Techniques: Injection, Derivative Assay, Microscopy, Transplantation Assay, Confocal Microscopy, Control, Double Staining, Staining

Journal: The Journal of Clinical Investigation

Article Title: Long-term corneal recovery by simultaneous delivery of hPSC-derived corneal endothelial precursors and nicotinamide

doi: 10.1172/JCI146658

Figure Lengend Snippet: ( A ) Corneal transparency and anterior segment OCT images of monkeys with CEP injection with (M2–M5) or without NAM treatment (M1) 3 months, 6 months, and 36 months after operation. Normal monkey (Norm) was used as the control. ( B ) Corneal thicknesses were measured by pachymeter at preoperation (Pre), 3 days, 2 weeks, 4 weeks, 8 weeks, and 6 months after transplantation. The dashed line shows normal corneal thickness. ( C ) Corneal confocal microscopy observation 3 months, 6 months, and 36 months after transplantation. No images of M1 without NAM treatment were obtained due to corneal edema. ( D ) Schematic representation of the scrape area in the half cornea of M4 with CEP and NAM treatment 36 months after operation. The 6 mm diameter scraped region was divided into 3 areas: Φ ≤ 2 mm, 2 mm ≤ Φ ≤ 4 mm, and 4 mm ≤ Φ ≤ 6 mm. Immunostaining of human cell–specific marker SC-121 and corneal endothelial marker ZO-1 in the different areas of the monkey corneal endothelium is shown. Nuclei were stained with DAPI. Scale bar: 50 μm. ( E ) Double staining of human cell–specific marker SC-121 and corneal endothelial markers ZO-1 and ATP1A1 in corneal endothelium of M4 36 months after transplantation. Nuclei were stained with DAPI. Scale bar: 50 μm.

Article Snippet: The cells were viewed under bright field microscopy (Echo Laboratories).

Techniques: Injection, Control, Transplantation Assay, Confocal Microscopy, Immunostaining, Marker, Staining, Double Staining