Clinical outcome of autologous cultivated limbal epithelium transplantation

Sangwan, Virender S; Matalia, Himanshu; Vemuganti, Geeta K.; Anjum, Fatima; Ifthekar, Ghazala; Singh, Shashi; Nutheti, Rishita; Rao, Gullapalli N

doi:10.4103/0301-4738.21611

Cited by 142 publications

(82 citation statements)

References 19 publications

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“…In cases of unilateral limbal stem cell deficiency (LSCD), a small piece of limbal biopsy approximately 2 mm in size is harvested from the healthy fellow eye and used as the stem cell source for in vitro culture and expansion of limbal stem cells in autologous cultured limbal epithelial transplantations (CLETs) [4]. The two popular culture methods used for the CLET procedure are direct explant cultures (ECs) on complex biological substrates such as human amniotic membrane (hAM) [5][6][7][8] and the culture of enzymatically dissociated cell suspensions on mitotically inactivated murine NIH3T3 feeders [4,9]. Both culture methods result in expansion of limbal stem cells in vitro and generate a sheet of epithelial monolayer that can be used to cover and regenerate the entire corneal surface of the affected eyes [10,11].…”

Section: Introductionmentioning

confidence: 99%

Spatial Distribution of Niche and Stem Cells in Ex Vivo Human Limbal Cultures

Mariappan

Kacham

Purushotham

et al. 2014

Stem Cells Translational Medicine

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Stem cells at the limbus mediate corneal epithelial regeneration and regulate normal tissue homeostasis. Ex vivo cultured limbal epithelial transplantations are being widely practiced in the treatment of limbal stem cell deficiency. In this report, we examined whether the limbal niche cells that nurture and regulate epithelial stem cells coexist in ex vivo limbal cultures. We also compared the inherent differences between explant and suspension culture systems in terms of spatial distribution of niche cells and their effect on epithelial stem cell proliferation, migration, and differentiation in vitro. We report that the stem cell content of both culture systems was similar, explaining the comparable clinical outcomes reported using these two methods. We also showed that the niche cells get expanded in culture and the nestin-positive cells migrate at the leading edges to direct epithelial cell migration in suspension cultures, whereas they are limited to the intact niche in explant cultures. We provide evidence that C/EBPd-positive, p15-positive, and quiescent, label-retaining, early activated stem cells migrate at the leading edges to regulate epithelial cell proliferation in explant cultures, and this position effect is lost in early suspension cultures. However, in confluent suspension cultures, the stem cells and niche cells interact with each another, migrate in spiraling patterns, and self-organize to form three-dimensional niche-like compartments resembling the limbal crypts and thereby reestablish the position effect. These 3D-sphere clusters are enriched with nestin-, vimentin-, S100-, and p27-positive niche cells and p15-, p21-, p63a-, C/EBPd-, ABCG2-, and Pax6-positive quiescent epithelial stem cells. STEM CELLS

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Section: Introductionmentioning

confidence: 99%

Spatial Distribution of Niche and Stem Cells in Ex Vivo Human Limbal Cultures

Mariappan

Kacham

Purushotham

et al. 2014

Stem Cells Translational Medicine

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“…Since the corneal surface is spherical, the large-diameter filter paper cannot fully adhere to the 260 degree corneal limbus,2 moderate and severe alkaline burn injury model prepared with different filter papers at a diameter less than 12 mm in the center of corneal tissues 13) . At present, limbal stem cell transplantation applied in clinical contexts mainly includes autologous or allogenic corneal limbal tissues and in vitro cultured limbal stem cells [14][15][16][17] . It is suitable to adopt a small amount of healthy corneal limbal tissue to culture a large area of corneal epithelial for in vitro culture 18,19) .…”

Section: Discussionmentioning

confidence: 99%

Tissue-Engineered Corneal Epithelium Transplantation for Repair of Corneal Alkali Burn

Lü

Tang

et al. 2012

J. Hard Tissue Biology.

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The tissue-engineered corneal epithelium can be constructed with cultured limbal stem cells from the cornea of healthy rabbits cornea cultured in vitro, and its transplantation may promote the repair and healing of corneal alkaline burn. The aim of this study was to probe the effect and opportunity of treating corneal alkali burn with a tissue-engineered corneal epithelium transplantation. The tissue-engineered corneal epithelium was prepared in 37 rabbits with limbal stem cells cultured in vitro, for transplantation in a double eye corneal alkali burn model. Autologous or allogenic transplantation was performed at early time points (1, 3, 6, 9 days) or metaphase (14 days) following alkaline burning. 1 week following the burn, large corneal epithelial desquamation occurred, with 72% of the rabbits having epithelial desquamations or corneal ulcers at 2 weeks, continuing to 4 weeks post burn. Only 25% of the rabbits in the transplantation group had these effects 4 weeks post-treatment. Cell infiltration and vascularization of corneal stroma was observed in the metaphase transplantation group, not the early transplantation groups; Within 4 weeks, immunological rejection induced by allogenic transplantation was not greater than that of autologous transplantation. Cultivated autologous or allogenic tissue-engineered corneal epithelium transplantation restored ocular surface integrity, and the effect of early transplantation surpasses that of metaphase.

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“…In this study ex vivo cultured limbal and conjunctival epithelial cells were grown on AM. In 2006, Sangwan et al published a more detailed report on clinical outcomes for 88 eyes of 86 patients (Sangwan et al, 2006). In 2001, Rama et al reported treatment outcomes for 18 eyes of 18 patients (Rama et al, 2001).…”

Section: Clinical Trials and Outcomes: Present And Futurementioning

confidence: 99%

“…Since 1997's landmark report, a variety of culture techniques have been developed to produce contiguous epithelial cell sheets for transplantation. These techniques can be broadly defined as either explant culture in which cells migrate out from limbal tissue attached to a surface (Grueterich et al, 2002b, Koizumi et al, 2001a, Koizumi et al, 2001b, Sangwan et al, 2006 or suspension culture in which cells are released from enzymatically digested extracellular matrix before culture (Daya et al, 2005, Pellegrini et al, 1997. The aforementioned methods have been used in studies to culture limbal epithelial cells successfully, on either a growth-arrested 3T3 fibroblast feeder layer or an amniotic membrane (AM), with varying results (Kim et al, 2004, Zito-Abbad et al, 2006.…”

Section: Ex Vivo Expansion Of Corneal Limbal Stem Cellsmentioning

confidence: 99%