To describe a novel surgical technique of in-vivo limbal stem cell cultivation for reconstruction of corneal epithelium in unilateral total limbal stem cell deficiency(LSCD).

: patients with chronic total unilateral LSCD due to a chemical injury were included. one small limbal block (2x1mm) harvested from contralateral healthy eye and sutured on the amniotic membrane on the diseased cornea. Amniotic membrane extract was used until the total corneal epithelialization was achieved. The eyes underwent subsequent optical penetrating keratoplasty and the corneal button analyzed for histopathologic exam and molecular analysis for detecting limbal stem cells.

In all 4 eyes total epithelialization of corneal surface was achieved in 2 weeks with regression of corneal conjunctivalization and vascularisation occurred between 3 to 4 weeks after surgery and was maintained at mean follow up of 6 months without any complication for donor and recipient eyes. Histopathologic exam and molecular analysis of corneal epithelium showed limbal Stem CellS over the cornea button.

In-vivo cultivation of limbal stem cells is an effective procedure for ocular surface reconstruction in patients with unilateral total LSCD . 6 months after surgery, all cornea were much clearer with regression of vascularisation . It’s long term effect should be confirmed in studies with longer follow up .