The Application of Cultured Cornea Stem Cells in Patients Suffering From Corneal Stem Cell Insufficiency

Study Overview
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Brief Summary
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Name of the project：The application of cornea stem cells cultured on amniotic membrane in patients suffering from corneal stem cell insufficiency Purpose: To develop the technique of using ex vivo expansion of autologous limbal stem cell cultured on amniotic membrane for treatment of patients suffering from limbal stem cell insufficiency

Detailed Description
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Materials and Methods: Patients suffering from unilateral limbal stem cell insufficiency will be chosen. Make sure those volunteers understand the benefits and risk factors of the operation. Prepare human amniotic membrane, and culture the membrane at37°C，5% CO2，95% room air. One month before transplantation, take 1-2mm2 of limbal tissues from the healthy eye, and culture the limbal tissue on amniotic membrane with 1.5 ml supplemental hormonal epithelial medium (in 5%的 fetal calf serum) Keep culturing the cells in Dulbecco's modified Eagle's medium/Ham's F12 (1:1 ratio), added with 5 % dimethyl sulfoxide, 2 mg/ml human epidermal growth factor ,5 ug/ml insulin, 0.5 ug/ml hydrocortisone, 50 ug/ml gentamicin, 1.25 ug/ml amphotericin B, 5% autoserum。Two to three weeks later, transplantation will be performed after the cells reaches 2-3 cm2 Remove the abnormal surface tissue on the lesion cornea, transplant the amniotic membrane with cultured limbal stem cells on the denuded cornea. Cover with contact lens after operation, and apply topical antibiotics and steroids. Postoperatively, observe the patients for one week. After discharge, follow up the patients one week, two week, one month, two months, three months, six months and one year later. Pictures will be taken and the extent of corneal epithelium regeneration, visual acuity, and complications will be recorded during follow-up visits. The investigators plan to collect 20 patients from 1 July,2007 to 30, June, 2010

Official Title
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Phase 1 Study of Cornea Stem Cells Cultured on Amniotic Membrane Transplanted on Patients Suffering From Corneal Stem Cell Insufficiency

Conditions
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Unilateral Limbal Stem Cell Insufficiency

Intervention / Treatment
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* Procedure: Transplant of cultured limbal stem cells on cornea


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: unilateral limbus defect larger than 180 degrees repeated corneal epithelial ulcer, loss of visual acuity or neovascularization due to limbal stem cell insufficiency no improvement after medical intervention for more than 6 months the patient is willing to receive surgery and postoperative follow up Exclusion Criteria: bilateral limbus defect limbal stem cell insufficiency does not induce repeated corneal epithelial ulcer, loss of visual acuity or neovascularization corneal neovascularization deeper than 1/2 depth of surface matrix peripheral surface matrix is less than 200um thick no corneal reflex was induced by fiber less than 5mm length measured by Cocet-Bonnet esthesiometer improvement under medication perisurgical follow up less than 6 months unable to be long term followed up postoperatively glaucoma patients underwent glaucoma surgery, under medication, IOP > 21mmHg glaucoma patients underwent glaucoma surgery, under medication, IOP < 21mmHg, optic disc depression > 90% severe incomplete eyelid closure severe dye eye syndrome, Schirmer test 5 min < 3 mm contralateral eye limbus tissue cannot be obtained, or unable to be cultured pregnant
Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Non-Randomized
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: 1<br>Patients suffering from unilateral limbal stem cell insufficiency. Remove the abnormal surface tissue on the lesion cornea, transplant the amniotic membrane with cultured limbal stem cells on the denuded cornea. Cover with contact lens after operation, and apply topical antibiotics and steroids. | Procedure: Transplant of cultured limbal stem cells on cornea<br>* Remove the abnormal surface tissue on the lesion cornea, transplant the amniotic membrane with cultured limbal stem cells on the denuded cornea. Cover with contact lens after operation, and apply topical antibiotics and steroids.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The extent of corneal epithelium regeneration |  | one week, two week, one month, two months, three months, six months and one year later | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| visual acuity |  | one week, two week, one month, two months, three months, six months and one year later | 
| complications |  | one week, two week, one month, two months, three months, six months and one year later | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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limbal stem cell, amniotic membrane, ex vivo expansion of autologous limbal stem cell, unilateral limbal stem cell insufficiency