Human corneal endothelial cell transplantation in a human ex vivo model

Sanjay V. Patel, Lori A. Bachman, Cheryl R. Hann, Cindy K. Bahler, Michael P Fautsch

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

PURPOSE. To determine the effects of incorporating superpara- magnetic microspheres (SPMs) into cultured human corneal endothelial cells (HCECs) and to describe preliminary experi- ments of HCEC transplantation, facilitated by SPMs and an external magnetic field, in a human anterior segment ex vivo model. METHODS. HCECs were cultured as monolayers and incorpo- rated with magnetite oxide SPMs (900, 300, and 100 nm) at different concentrations. Cell viability, migration toward a magnetic field, and light transmittance were measured after incorporation of the SPMs. HCEC transplantation into the eyes of human recipients was investigated by subjecting anterior segments in organ culture to an external magnetic field. Light and electron microscopy were used to assess HCEC attachment to corneal stroma. RESULTS. SPMs were incorporated into the cytoplasm of HCECs after overnight incubation. None of the SPMs affected the short-term viability of cultured HCECs (P 0.14, n 6) or their light transmittance (P 0.06, n 5), although there was a trend toward decreased transmittance with the higher concentration of 900-nm SPMs. Cell migration toward a magnetic field was higher for HCECs with incorporated SPMs than for HCECs without SPMs (P 0.01, n 6), with dose-response relationships evident for the 300- and 100-nm SPMs. SPMs facilitated the attachment of HCECs to the corneal stroma in the human anterior segment model with minimal change in intracameral (intraocular) pressure. CONCLUSIONS. SPMs facilitate migration of HCECs toward a magnetic source and attachment of cells to the corneal stroma without affecting cell viability or light transmittance. The human anterior segment model can be used to study HCEC transplantation.

Original languageEnglish (US)
Pages (from-to)2123-2131
Number of pages9
JournalInvestigative Ophthalmology and Visual Science
Volume50
Issue number5
DOIs
StatePublished - 2009

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Cell Transplantation
Endothelial Cells
Microspheres
Magnetic Fields
Corneal Stroma
Light
Cell Movement
Cell Survival
Ferrosoferric Oxide
Organ Culture Techniques
Intraocular Pressure
Oxides

ASJC Scopus subject areas

  • Medicine(all)
  • Ophthalmology
  • Cellular and Molecular Neuroscience
  • Sensory Systems

Cite this

Human corneal endothelial cell transplantation in a human ex vivo model. / Patel, Sanjay V.; Bachman, Lori A.; Hann, Cheryl R.; Bahler, Cindy K.; Fautsch, Michael P.

In: Investigative Ophthalmology and Visual Science, Vol. 50, No. 5, 2009, p. 2123-2131.

Research output: Contribution to journalArticle

Patel, Sanjay V. ; Bachman, Lori A. ; Hann, Cheryl R. ; Bahler, Cindy K. ; Fautsch, Michael P. / Human corneal endothelial cell transplantation in a human ex vivo model. In: Investigative Ophthalmology and Visual Science. 2009 ; Vol. 50, No. 5. pp. 2123-2131.
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N2 - PURPOSE. To determine the effects of incorporating superpara- magnetic microspheres (SPMs) into cultured human corneal endothelial cells (HCECs) and to describe preliminary experi- ments of HCEC transplantation, facilitated by SPMs and an external magnetic field, in a human anterior segment ex vivo model. METHODS. HCECs were cultured as monolayers and incorpo- rated with magnetite oxide SPMs (900, 300, and 100 nm) at different concentrations. Cell viability, migration toward a magnetic field, and light transmittance were measured after incorporation of the SPMs. HCEC transplantation into the eyes of human recipients was investigated by subjecting anterior segments in organ culture to an external magnetic field. Light and electron microscopy were used to assess HCEC attachment to corneal stroma. RESULTS. SPMs were incorporated into the cytoplasm of HCECs after overnight incubation. None of the SPMs affected the short-term viability of cultured HCECs (P 0.14, n 6) or their light transmittance (P 0.06, n 5), although there was a trend toward decreased transmittance with the higher concentration of 900-nm SPMs. Cell migration toward a magnetic field was higher for HCECs with incorporated SPMs than for HCECs without SPMs (P 0.01, n 6), with dose-response relationships evident for the 300- and 100-nm SPMs. SPMs facilitated the attachment of HCECs to the corneal stroma in the human anterior segment model with minimal change in intracameral (intraocular) pressure. CONCLUSIONS. SPMs facilitate migration of HCECs toward a magnetic source and attachment of cells to the corneal stroma without affecting cell viability or light transmittance. The human anterior segment model can be used to study HCEC transplantation.

AB - PURPOSE. To determine the effects of incorporating superpara- magnetic microspheres (SPMs) into cultured human corneal endothelial cells (HCECs) and to describe preliminary experi- ments of HCEC transplantation, facilitated by SPMs and an external magnetic field, in a human anterior segment ex vivo model. METHODS. HCECs were cultured as monolayers and incorpo- rated with magnetite oxide SPMs (900, 300, and 100 nm) at different concentrations. Cell viability, migration toward a magnetic field, and light transmittance were measured after incorporation of the SPMs. HCEC transplantation into the eyes of human recipients was investigated by subjecting anterior segments in organ culture to an external magnetic field. Light and electron microscopy were used to assess HCEC attachment to corneal stroma. RESULTS. SPMs were incorporated into the cytoplasm of HCECs after overnight incubation. None of the SPMs affected the short-term viability of cultured HCECs (P 0.14, n 6) or their light transmittance (P 0.06, n 5), although there was a trend toward decreased transmittance with the higher concentration of 900-nm SPMs. Cell migration toward a magnetic field was higher for HCECs with incorporated SPMs than for HCECs without SPMs (P 0.01, n 6), with dose-response relationships evident for the 300- and 100-nm SPMs. SPMs facilitated the attachment of HCECs to the corneal stroma in the human anterior segment model with minimal change in intracameral (intraocular) pressure. CONCLUSIONS. SPMs facilitate migration of HCECs toward a magnetic source and attachment of cells to the corneal stroma without affecting cell viability or light transmittance. The human anterior segment model can be used to study HCEC transplantation.

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