June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Isolation of Human Corneal Endothelial Cells and Potential for Immediate Transplantation
Author Affiliations & Notes
  • Behnam Rabiee
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Rebecca Jung
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Ilham Putra
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Khandaker N Anwar
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Yasmin Rassouli
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Xiang Shen
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Ghasem Yazdanpanah
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Sayena Jabbehdari
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Neda Afsharkhamseh
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Mark Rosenblatt
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Mahmood Ghassemi
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Ali R Djalilian
    Illinois Eye and Ear Infirmary, UIC, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Behnam Rabiee, None; Rebecca Jung, None; Ilham Putra, None; Khandaker Anwar, None; Yasmin Rassouli, None; Xiang Shen, None; Ghasem Yazdanpanah, None; Sayena Jabbehdari, None; Neda Afsharkhamseh, None; Mark Rosenblatt, None; Mahmood Ghassemi, None; Ali Djalilian, None
  • Footnotes
    Support  This work was supported by R01 EY024349 (ARD) and Core grant EY01792 from NEI/NIH, unrestricted grant to the department from Research to Prevent Blindness, and Eversight.
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 3598. doi:
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      Behnam Rabiee, Rebecca Jung, Ilham Putra, Khandaker N Anwar, Yasmin Rassouli, Xiang Shen, Ghasem Yazdanpanah, Sayena Jabbehdari, Neda Afsharkhamseh, Mark Rosenblatt, Mahmood Ghassemi, Ali R Djalilian; Isolation of Human Corneal Endothelial Cells and Potential for Immediate Transplantation. Invest. Ophthalmol. Vis. Sci. 2020;61(7):3598.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Pioneering studies have shown that cultivated human corneal endothelial cells (HCEC) can be used to restore a functional human corneal endothelium in disease states. Inspired by these studies, we conducted this project to determine the feasibility of isolating HCECs for potential immediate transplantation (without ex-vivo propagation).

Methods : Donor corneas prepared for Descemet Membrane Endothelial Keratoplasty (DMEK) were digested in different concentrations of collagenase, in different media for different durations. Following digestion, the centrifugation process was optimized with different buffers, speeds, and time points. Isolated cells were labeled with a live cell tracker. These cells were then placed on the endothelial surface of donor corneas after scraping the endothelial cells ex vivo. The isolated cells were also cultured in vitro to monitor their morphology and characteristics.

Results : The optimized digestion condition was found to be 2 hours at 37°C in a medium consisting of OptiMEM reduced serum media, 8% human serum, 1mg/mL collagenase, 200µg/mL calcium chloride, and 50µg/mL gentamycin. Optimized cellular recovery (80±2%) and viability (89±1%) was obtained at 4000g in 10 minutes after adding 1:1 human serum to the digestion media containing the isolated cells. The isolated cells were viable and successfully attached to the endothelial surface of donor corneas ex vivo after 3 hours and remained viable and attached after 9 days. In vitro culture of the isolated cells resulted in characteristic morphology of hexagonal endothelial cells after 14 days.

Conclusions : These findings show that human corneal endothelial cells can be isolated with high viability which could potentially be used for endothelial cell transplantation immediately. This approach limits the in vitro manipulation of the cells which may be advantageous from a safety and regulatory standpoint.

This is a 2020 ARVO Annual Meeting abstract.

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