1. Himanshu Matalia, MD1,2 Arokiaraj Vincent, MSc2 D Kamesh, MSc2
Early results of ex vivo cultivated autologous limbal epithelium transplantation for limbal stem cell deficiency
Himanshu Matalia, MD1,2
Arokiaraj Vincent, MSc2
D Kamesh, MSc2
Cornea & Refractive Services, Narayana Nethralaya
Narayana Nethralaya Stem Cell Research Laboratory
Narayana Nethralaya
Narayana Health City
Super Specialty Eye Hospital &
Post Graduate Institute of Ophthalmology
Bangalore - INDIA
Authors have no financial interest
Poster 610

2. Limbal Stem Cell Defiency (LSCD)
Clinical Features of LSCD
Loss of palisades of Vogt
Kinoshita S et al JpJ Clin Ophth 1986
Tseng SC et al OCNA 1990
Conjunctivalization
Stippled appearance with fluorescein
Dua HS Am J Ophthalmol 1990
Huang AJ IOVS 1991
Ch. Inflammation
Recurrent epi. break down

3. Algorithm for LSCD Unilateral Disease Bilateral Disease
Establish Diagnosis of Stem cells deficiency (Clinical features & Impression Cytology)
Unilateral Disease
Bilateral Disease
Focal or
Partial
Total
Deficiency
KLAL
Lr-CLAL
Cultured live-related limbal allograft
AMT
CLAG
Debridment
Cadaveric kerato-limbal allograft (KLAL)
Live-related conjunctivo limbal allograft (Lr-CLAL)
Cultured limbal auto graft (best option )
Outcome not very encouraging

4. Cultivated Autologous Limbal Epithelium Transplantation (CALET)
Most recent technique
Best for unilateral LSCD
Autologous cells, no immune suppression required
No risk at donor site
Repeatable
Limitations
Availability of the tissue culture facility
Long-term outcome awaited
Pellegrini et al Lancet 1997; 349:
Tsai RJ, Li LM, Chen JK. N Engl J Med 2000;343(2):86-93
Schwab et al Cornea 2000;19(4):421-6
Koizumi et al IOVS 2002;43(7):

5. Purpose
To describe the early results of our technique of ex vivo cultivated autologous limbal epithelial transplantation (CALET) for limbal stem cell deficiency (LSCD).

6. Methods Prospective, non-randomized interventional case series
14 eyes of 14 consecutive patient with LSCD underwent CALET from October 2008-October 2009.
Primary outcome measures:
Successful ocular surface reconstruction: stable ocular surface
Best corrected visual acuity (BCVA)
Secondary outcome measures:
Complications

7. Culture technique Limbal biopsy Amniotic membrane Unique from others*
Normal limbal epithelium from contra lateral normal eye or normal area of same eye
2 x 2 mm (One clock hour) of normal limbal with cornea epithelium
Amniotic membrane
De-epithelialized, processed & preserved amniotic membrane used as substrate / carrier
Unique from others*
Monolayer only
No air lifting, no fibroblast feeder
Culture duration: days
Human Corneal Epithelium medium
*Sangwan VS, Matalia HP et al Arch Ophthalmol 2005

8. Surgical technique
One drop 1:1000 epinephrine on pannus to reduce bleeding (chemical cauterization)
Pannus resection done
Conjunctival peri-ectomy (2 mm) was done
Cauterization of bleeders done
Ex vivo cultivated autologous limbal epithelium grown over human amnotic membrane (HAM) was spread over the ocular surface
HAM was secured to the eye with fibrin glue (Tisseal) and 8-0 Vicryl
Bandage contact lens applied at the end

9. Results From July 2008 14 eyes of 14 patients underwent CALET
Female: Male -
Etiology of LSCD:
Chemical injury 10 eyes (72%)
8 of these 10 were lime injury
7 eyes (50%): Previous history of AMT
Follow up: Mean 9 months
Mean duration of culture: 174.40 days

10. Results Clinical Outcome Visual acuity improvement Complications
Preop
Clinical Outcome
Success: 12 (85.7), 95% CI
Failure: 2 (14.3%), 95%CI:
Visual acuity improvement
Preop visual acuity: 0.20  0.38
Postop visual acuity: 0.38 0.41 (p=0.018)
Complications
Microbial keratitis: 2
Repeat transplants: 2
Repeat limbal biopsy: 2
HM+ PR accurate
Postop
20/4020/25P

11. Results Visual acuity improvement Preop visual acuity: 20/100
Postop visual acuity: 20/50 (p=0.018)

12. Conclusions
Cultivated autologous limbal epithelial transplantation (CALET) could successfully restore the ocular surface in cases with LSCD.