Human corneal endothelial cell transplantation in a human ex vivo model.
OBJECTIVE
To determine the effects of incorporating superparamagnetic microspheres (SPMs) into cultured human corneal endothelial cells (HCECs) and to describe preliminary experiments 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 incorporated 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 < or = 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.
Patel
Sanjay V
SV
Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota 55905, USA. patel.sanjay@mayo.edu
Bachman
Lori A
LA
Hann
Cheryl R
CR
Bahler
Cindy K
CK
Fautsch
Michael P
MP
D000293:Adolescent; D000328:Adult; D000869:Anterior Eye Segment / Q000601:surgery*; D002452:Cell Count; D002465:Cell Movement / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D017690:Cell Transplantation; D002478:Cells, Cultured; D004728:Endothelium, Corneal / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure; D052203:Ferrosoferric Oxide / Q000378:metabolism; D006801:Humans; D008280:Magnetics; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D008863:Microspheres; D008875:Middle Aged; D008954:Models, Biological; D009924:Organ Culture Techniques
D052203:Ferrosoferric Oxide