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Stem Cell Therapy for Corneal Epithelium Regeneration following Good Manufacturing and Clinical Procedures.

Ramírez BE, Sánchez A, Herreras JM, Fernández I, García-Sancho J, Nieto-Miguel T, Calonge M - Biomed Res Int (2015)

Bottom Line: All clinical parameters improved substantially.By IVCM, 80% of cases improved in epithelial status.These results confirm that CLET is a valid therapy for ocular surface failure.

View Article: PubMed Central - PubMed

Affiliation: Institute of Applied Ophthalmobiology (IOBA), University of Valladolid, Campus Universitario Miguel Delibes, Paseo de Belén 17, 47011 Valladolid, Spain.

ABSTRACT

Objective: To evaluate outcomes of cultivated limbal epithelial transplantation (CLET) for management of ocular surface failure due to limbal stem cell deficiency (LSCD).

Design: Prospective, noncomparative, interventional case series and extensive comparison with recent similar studies.

Participants: Twenty eyes with LSCD underwent CLET (11 autologous; 9 allogeneic) and were followed up for 3 years. Etiologies were divided into 3 prognostic categories: Group 1, chemical injuries (7 eyes); Group 2, immune-based inflammation (4 eyes); and Group 3, noninflammatory diseases (9 eyes). Intervention. Autologous and allogeneic limbal epithelial cells were cultivated on amniotic membranes and transplanted. Evaluations were based on clinical parameters, survival analysis, and in vivo confocal microscopy (IVCM). European Union Tissues/Cells Directive and good manufacturing procedures were followed.

Main outcome measures: Improved clinical parameters, absence of epithelial defects, and improved central corneal epithelial phenotype.

Results: Success rate was 80% at 1-2 years and 75% at 3 years. Autografts and allografts had similar survival. Success rate was significantly lower in prognostic Group 1 (42.9%) than in Groups 2-3 (100% each). All clinical parameters improved substantially. By IVCM, 80% of cases improved in epithelial status.

Conclusions: CLET improved corneal epithelium quality, with subsequent improvement in symptoms, quality of life, and vision. These results confirm that CLET is a valid therapy for ocular surface failure.

No MeSH data available.


Related in: MedlinePlus

Limbal biopsy, limbal epithelial cultivation, and cultivated limbal epithelial transplantation (CLET). (a) Healthy donor eye 24 hr. after a 2 × 2 mm limbal biopsy; (b) the biopsy tissue was placed in an Eppendorf tube with culture medium (b1); (c) the biopsy was processed in a good manufacturing practice-cell processing unit within the next 4 hr. and for the next 4-5 weeks; and (d) the explant was placed on denuded human amniotic membrane, as viewed by contrast phase microscopy. Limbal epithelial cells began outgrowth from the explant at 1-2 weeks (d1). The explant was then removed, and the outgrowth was maintained until reaching confluence at which time it contained approximately 250,000 cells. The cell product was then sent to the medical center for CLET (d2). (e) Superficial keratectomy in the diseased contralateral eye (Case 1); (e1) human amniotic membrane with epithelial limbal cells confluent on top is removed from culture dish; (e2) the complex of amniotic membrane-limbal stem cells is placed on top of the previously denuded corneal and sclerolimbal surface; the amniotic membrane limit is observed (black arrow) with cells facing down and sutured (white arrows). A scleral lens is then applied.
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fig1: Limbal biopsy, limbal epithelial cultivation, and cultivated limbal epithelial transplantation (CLET). (a) Healthy donor eye 24 hr. after a 2 × 2 mm limbal biopsy; (b) the biopsy tissue was placed in an Eppendorf tube with culture medium (b1); (c) the biopsy was processed in a good manufacturing practice-cell processing unit within the next 4 hr. and for the next 4-5 weeks; and (d) the explant was placed on denuded human amniotic membrane, as viewed by contrast phase microscopy. Limbal epithelial cells began outgrowth from the explant at 1-2 weeks (d1). The explant was then removed, and the outgrowth was maintained until reaching confluence at which time it contained approximately 250,000 cells. The cell product was then sent to the medical center for CLET (d2). (e) Superficial keratectomy in the diseased contralateral eye (Case 1); (e1) human amniotic membrane with epithelial limbal cells confluent on top is removed from culture dish; (e2) the complex of amniotic membrane-limbal stem cells is placed on top of the previously denuded corneal and sclerolimbal surface; the amniotic membrane limit is observed (black arrow) with cells facing down and sutured (white arrows). A scleral lens is then applied.

Mentions: Donor human amniotic membranes and cadaveric limbal rings came from a registered and accredited tissue bank (Blood and Tissue Community Center, Oviedo, Asturias, Spain) and were screened for transmittable diseases as described above. Allogeneic limbal rings were obtained within 7 days of death from corneal donors who were under 60 years old. Autologous limbal biopsies were marked by the surgeon with a stitch in the upper right corner to ensure consistency when plating the explant to ensure the same way up; they were sent from the operating room to the cell production unit at 4–10°C in culture medium (Figure 1).


Stem Cell Therapy for Corneal Epithelium Regeneration following Good Manufacturing and Clinical Procedures.

Ramírez BE, Sánchez A, Herreras JM, Fernández I, García-Sancho J, Nieto-Miguel T, Calonge M - Biomed Res Int (2015)

Limbal biopsy, limbal epithelial cultivation, and cultivated limbal epithelial transplantation (CLET). (a) Healthy donor eye 24 hr. after a 2 × 2 mm limbal biopsy; (b) the biopsy tissue was placed in an Eppendorf tube with culture medium (b1); (c) the biopsy was processed in a good manufacturing practice-cell processing unit within the next 4 hr. and for the next 4-5 weeks; and (d) the explant was placed on denuded human amniotic membrane, as viewed by contrast phase microscopy. Limbal epithelial cells began outgrowth from the explant at 1-2 weeks (d1). The explant was then removed, and the outgrowth was maintained until reaching confluence at which time it contained approximately 250,000 cells. The cell product was then sent to the medical center for CLET (d2). (e) Superficial keratectomy in the diseased contralateral eye (Case 1); (e1) human amniotic membrane with epithelial limbal cells confluent on top is removed from culture dish; (e2) the complex of amniotic membrane-limbal stem cells is placed on top of the previously denuded corneal and sclerolimbal surface; the amniotic membrane limit is observed (black arrow) with cells facing down and sutured (white arrows). A scleral lens is then applied.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4588357&req=5

fig1: Limbal biopsy, limbal epithelial cultivation, and cultivated limbal epithelial transplantation (CLET). (a) Healthy donor eye 24 hr. after a 2 × 2 mm limbal biopsy; (b) the biopsy tissue was placed in an Eppendorf tube with culture medium (b1); (c) the biopsy was processed in a good manufacturing practice-cell processing unit within the next 4 hr. and for the next 4-5 weeks; and (d) the explant was placed on denuded human amniotic membrane, as viewed by contrast phase microscopy. Limbal epithelial cells began outgrowth from the explant at 1-2 weeks (d1). The explant was then removed, and the outgrowth was maintained until reaching confluence at which time it contained approximately 250,000 cells. The cell product was then sent to the medical center for CLET (d2). (e) Superficial keratectomy in the diseased contralateral eye (Case 1); (e1) human amniotic membrane with epithelial limbal cells confluent on top is removed from culture dish; (e2) the complex of amniotic membrane-limbal stem cells is placed on top of the previously denuded corneal and sclerolimbal surface; the amniotic membrane limit is observed (black arrow) with cells facing down and sutured (white arrows). A scleral lens is then applied.
Mentions: Donor human amniotic membranes and cadaveric limbal rings came from a registered and accredited tissue bank (Blood and Tissue Community Center, Oviedo, Asturias, Spain) and were screened for transmittable diseases as described above. Allogeneic limbal rings were obtained within 7 days of death from corneal donors who were under 60 years old. Autologous limbal biopsies were marked by the surgeon with a stitch in the upper right corner to ensure consistency when plating the explant to ensure the same way up; they were sent from the operating room to the cell production unit at 4–10°C in culture medium (Figure 1).

Bottom Line: All clinical parameters improved substantially.By IVCM, 80% of cases improved in epithelial status.These results confirm that CLET is a valid therapy for ocular surface failure.

View Article: PubMed Central - PubMed

Affiliation: Institute of Applied Ophthalmobiology (IOBA), University of Valladolid, Campus Universitario Miguel Delibes, Paseo de Belén 17, 47011 Valladolid, Spain.

ABSTRACT

Objective: To evaluate outcomes of cultivated limbal epithelial transplantation (CLET) for management of ocular surface failure due to limbal stem cell deficiency (LSCD).

Design: Prospective, noncomparative, interventional case series and extensive comparison with recent similar studies.

Participants: Twenty eyes with LSCD underwent CLET (11 autologous; 9 allogeneic) and were followed up for 3 years. Etiologies were divided into 3 prognostic categories: Group 1, chemical injuries (7 eyes); Group 2, immune-based inflammation (4 eyes); and Group 3, noninflammatory diseases (9 eyes). Intervention. Autologous and allogeneic limbal epithelial cells were cultivated on amniotic membranes and transplanted. Evaluations were based on clinical parameters, survival analysis, and in vivo confocal microscopy (IVCM). European Union Tissues/Cells Directive and good manufacturing procedures were followed.

Main outcome measures: Improved clinical parameters, absence of epithelial defects, and improved central corneal epithelial phenotype.

Results: Success rate was 80% at 1-2 years and 75% at 3 years. Autografts and allografts had similar survival. Success rate was significantly lower in prognostic Group 1 (42.9%) than in Groups 2-3 (100% each). All clinical parameters improved substantially. By IVCM, 80% of cases improved in epithelial status.

Conclusions: CLET improved corneal epithelium quality, with subsequent improvement in symptoms, quality of life, and vision. These results confirm that CLET is a valid therapy for ocular surface failure.

No MeSH data available.


Related in: MedlinePlus