Ex Vivo Expansion and Transplantation of Limbal Epithelial Stem Cells

Shortt, Alex J.; Secker, Genevieve A.; Rajan, Madhavan S.; Meligonis, George; Dart, John K.G.; Tuft, Stephen; Daniels, Julie T.

doi:10.1016/j.ophtha.2008.04.039

Cited by 164 publications

(136 citation statements)

References 42 publications

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“…The procedure was found to improve or to restore vision in a majority of the cases, and subsequent studies from other groups confirmed the potential of this new therapeutic approach [6][7][8][9][10][11][12].…”

Section: Introductionmentioning

confidence: 75%

“…For initiation of cultures, the epithelial cells may be dissociated by trypsin-EDTA [13][14][15], by dispase [16,17], or by a sequential combination of dispase and trypsin-EDTA with or without accompanying mechanical procedures [12]. In order to expand the dissociated cells ex vivo, however, most protocols include use of a medium supplemented with a mixture of hormones, growth factors, cholera toxin and also FBS.…”

Section: Introductionmentioning

confidence: 99%

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Trypsin for Dissociation of Limbal Cells for Engineering of Grafts May Induce DNA Strand Breaks in the Harvested Cells

2013

J Ocul Biol

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Section: Introductionmentioning

confidence: 75%

Section: Introductionmentioning

confidence: 99%

Trypsin for Dissociation of Limbal Cells for Engineering of Grafts May Induce DNA Strand Breaks in the Harvested Cells

2013

J Ocul Biol

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“…Briefly, human limbal tissue specimens were obtained from the normal fellow eye of patients following standard described techniques [11,12]. The harvested tissue pieces were placed on the surface of the de-epithelialized human amniotic membrane and human corneal epithelium growth medium [8] was added.…”

Section: Methodsmentioning

confidence: 99%

Phenotypic Evaluation of Severely Damaged Ocular Surface after Reconstruction by Cultured Limbal Epithelial Cell Transplantation

Sharma

Tandon²,

Mohanty³

et al. 2013

Ophthalmic Res

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Objective: To report the clinical outcome of autologous cultured limbal epithelial cell transplantation (CLECT) followed by deep anterior lamellar keratoplasty (DALK) in paediatric eyes and to correlate the clinical outcome with the phenotype of rejuvenated corneal epithelium. Methods: Four patients with total limbal stem cell deficiency (LSCD) underwent autologous CLECT. Cultivated cell sheets were transplanted onto the damaged ocular surface followed by DALK surgery. Excised corneal buttons were subjected to histopathological analysis. Data recorded included age, sex, laterality, nature of injury, follow-up period, severity of stem cell deficiency, visual acuity, Schirmer's test and impression cytology. Results: At a mean follow-up period of 19.5 ± 7.4 (range 9-26) months after CLECT, all 4 eyes showed epithelialized and clinically stabilized ocular surface. Manual DALK was performed in all 4 eyes, with a mean follow-up of 9.75 ± 4.5 (range 5-15) months. All eyes exhibited smooth and clear corneal epithelium with improved visual acuity. Excised corneal buttons demonstrated organized corneal epithelial morphology and showed expression of cornea-specific CK3/12 marker. Conclusion: Restoration of severely damaged ocular surface following chemical injury by using 2-stage meticulous approaches offers a new modality for the treatment of severe LSCD. Transplantation of cultivated autologous limbal epithelial cell sheet followed by DALK surgery can efficiently restore the corneal phenotype with improved vision.

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“…Druga metoda polega na trawieniu fragmentu rogówki z wykorzystaniem enzymów w celu oddzielenia komórek macierzystych od nabłonka, a następnie ich hodowli. Wykorzystuje się tu zwykle dyspazę, hydrolizującą kolagen, oraz trypsynę, rozdzielającą strukturę rąbka na pojedyncze komórki [18,69]. Brak jednak spójnych informacji, która technika jest wydajniejsza.…”

Section: Terapia Z Zastosowaniem Komórek Macierzystych Rąbka Rogówkiunclassified

Limbal epithelial stem cells in regeneration of corneal epithelium – clinical and molecular aspects

Sikora

Skubis

Sedlak

et al. 2018

Ann. Acad. Med. Siles.

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ST R ES ZCZ E NI EKomórki macierzyste rąbka rogówki (limbal epithelial stem cells -LESC) zlokalizowane są na granicy rogówki i spojówki. Ich funkcja polega na odbudowie rogówki poprzez zastępowanie uszkodzonych lub niefunkcjonalnych komórek. W niniejszej pracy zawarto informacje dotyczące charakterystyki LESC, opisano sposoby ich identyfikacji z wykorzystaniem markerów powierzchniowych oraz cech morfologicznych. W pracy zawarto również informacje dotyczące hodowli komórek w warunkach in vitro jako potencjalnego źródła komórek wykorzystywanych w terapii oraz poruszono zagadnienia dysfunkcji LESC spowodowanych niedoborem komórek macierzystych wywołanych różnymi czynnikami, a także przedstawiono stosowane obecnie metody leczenia LSCD (limbal stem cell deficiency). Standardowe leczenie niedoboru komórek macierzystych rąbka rogówki polega na farmakoterapii oraz transplantacji autologicznego rąbka rogówki bogatego w komórki macierzyste. W przypadku całkowitego niedoboru LESC konieczne jest wykonanie przeszczepów allogenicznych.

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Ex Vivo Expansion and Transplantation of Limbal Epithelial Stem Cells

Cited by 164 publications

References 42 publications

Trypsin for Dissociation of Limbal Cells for Engineering of Grafts May Induce DNA Strand Breaks in the Harvested Cells

Trypsin for Dissociation of Limbal Cells for Engineering of Grafts May Induce DNA Strand Breaks in the Harvested Cells

Phenotypic Evaluation of Severely Damaged Ocular Surface after Reconstruction by Cultured Limbal Epithelial Cell Transplantation

Limbal epithelial stem cells in regeneration of corneal epithelium – clinical and molecular aspects

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