Abstract
Purpose: :
To investigate the conservation of human anterior lens capsule containing grown limbal stem cells from cornea as a source of corneal tissue for corneal regeneration.
Materials and Methods: :
Anterior lens capsules from patients were removed during cataract surgery. Lens capsules were placed in distilled sterile water to remove any remaining epithelial cells adhered to the capsule. Lens capsules were plated on treated 35mm polystyrene tissue culture plates and limbal biopsies were collected in sterile tubes containing culture media DMEM/F12 and seeded onto lens capsules. At different time points autologous / heterologous colonized capsules were frozen in medium containing DMSO at –80ºC and then in liquid Nitrogen for at least two weeks. Frozen stored capsules were thawed and re–grown in normal culture medium.
Results: :
Anterior lens capsules colonized by limbal stem cells can be stored frozen in liquid nitrogen and then recovered to continue their normal cycle of division. We registered good proliferation starting on day 1, at 2 weeks and 1 month.
Conclusions: :
Corneal stem cells derived from limbal tissue grown over human anterior lens capsule, rich in collagen type IV, can be used as an available source of material for the treatment of limbal deficiencies or corneal degradation. The fact that this in vitro–generated tissue can be stored and maintained in liquid nitrogen until needed, makes it a very promising way of having a source of corneal tissue available at any time for patients needed.
Keywords: crystalline lens • regeneration • cornea: clinical science