Ex Vivo Preservation and Expansion of Human Limbal Epithelial Stem Cells on Amniotic Membrane for Treating Corneal Diseases with Total Limbal Stem Cell Deficiency

Tseng, Scheffer C.G.; Meller, Daniel; Anderson, David F.; Touhami, Amel; Pires, Renato T. F.; Grüterich, Martin; Solomon, Abraham; Espana, Edgar M.; Sandoval, Hugo; Ti, Seng-Ei; Goto, Eiki

doi:10.1007/978-1-4615-0717-8_192

Cited by 34 publications

(23 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…AM is a natural matrix substrate, which has been widely used for aiding the repair of corneal epithelium in experimental animals and in patients with corneal epithelial defects (Tseng et al, 1998;Meller and Tseng, 2000;Tseng et al, 2002;Ti et al, 2004). AM-based limbal explant culture has proved to be a good culture system to expand limbal epithelial cells to form limballike epithelia equivalents (Wang et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

The growth-promoting effect of KGF on limbal epithelial cells is mediated by upregulation of ΔNp63α through the p38 pathway

Cheng

Wang

Kao

et al. 2009

Journal of Cell Science

View full text Add to dashboard Cite

Corneal epithelial stem cells are thought to reside in the limbus, the transition zoon between cornea and conjunctiva. Keratinocyte growth factor (KGF) and hepatocyte growth factor (HGF) are two paracrine factors that regulate the proliferation, migration and differentiation of the limbal epithelial cells; however, the underlying mechanisms are still poorly understood. In an ex vivo limbal explant culture, we found that KGF is a more potent growth stimulator for the epithelial outgrowth than HGF. Immunofluorescence studies of the epithelial outgrowth from cells treated with HGF or KGF showed similar expression patterns of keratin-3 and keratin-14. Interestingly, p63 was highly expressed in KGF-treated limbal epithelial sheets but not in those treated with HGF. Kinase inhibitor studies showed that induction of ΔNp63α expression by KGF is mediated via the p38 pathway. The effect of KGF on limbal epithelial outgrowth was significantly reduced when endogenous ΔNp63α was suppressed, suggesting that KGF-induced limbal epithelial outgrowth is dependent on the expression of ΔNp63α. Our findings strongly suggest that limbal keratocytes regulate limbal epithelial cell growth and differentiation through a KGF paracrine loop, with ΔNp63α expression as one of the downstream targets.

show abstract

Section: Discussionmentioning

confidence: 99%

The growth-promoting effect of KGF on limbal epithelial cells is mediated by upregulation of ΔNp63α through the p38 pathway

Cheng

Wang

Kao

et al. 2009

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…Moreover, shortage of donor tissues is a serious current challenge. To avoid these problems, transplantations of cultivated corneal limbal epithelial cells have recently been reported [26][27][28][29][30][31]. In such tissue engineering approaches, development of new, effective methods for rapid identification and enrichment of corneal epithelial stem cells would be a considerable advantage since long-term, functional recovery of corneal epithelium is linked to graft constructs that retain viable stem cell populations [26,28,32].…”

Section: Discussionmentioning

confidence: 99%

Human limbal epithelium contains side population cells expressing the ATP-binding cassette transporter ABCG2

WATANABE¹

2004

FEBS Letters

View full text Add to dashboard Cite

“…A restoration of an intact corneal surface that is stable for more than 3 years would not be possible in patients with total limbal SC deficiency, without the transplantation of true SCs [41,42] . Our aim was to analyse the cell cycle kinetics, so we labelled the DNA with BrdU, and the following period resulted in the dilution 7 + population were determined as described in Materials. * p < 0.001 compared to respective cultures on AM.…”

Section: Discussionmentioning

confidence: 99%

Correlation of Cell Cycle Kinetics with p63 Expression in Human Limbal Epithelial Cells Expanded on Intact Human Amniotic Membrane

Galindo

Theiß²,

Pauklin

et al. 2008

Ophthalmic Res

Self Cite

View full text Add to dashboard Cite

Aim: To analyse the correlation of p63 expression and cell cycle kinetics of human limbal epithelial cells (HLECs) expanded on amniotic membrane (AM) or plastic. Methods: Primary HLECs were cultured either on cryopreserved intact AM or plastic dishes for 2 weeks. Cells were labelled with 5 μM 5-bromo-2′-deoxyuridine (BrdU) for 3 days, followed by an interval of either 7 or 14 days in BrdU-free medium. The expression of p63 and BrdU labelling was detected by immunocytochemistry. Results: More cells on AM (56%) than on plastic (24%) retained their BrdU label after the 7-day interval (p < 0.001). The difference was even more pronounced after 14 days (20 and 2%, p < 0.001). All BrdU-labelled cells were also p63 positive. 2.5-fold more cells on AM (54%) than on plastic (21%) were BrdU positive/p63 positive after 7 days (p < 0.001). It increased to a 9-fold difference after 14 days (p < 0.001). The BrdU label was lost more quickly than the p63 expression during the observation period, indicating that p63 expression was not confined to stem cells but existed also in transient amplifying cells. Conclusions: The combination of p63 expression and BrdU label retention is a better criterion to characterize stemness than either marker alone. AM as a culture substrate preserves stemness better than plastic.

show abstract

Ex Vivo Preservation and Expansion of Human Limbal Epithelial Stem Cells on Amniotic Membrane for Treating Corneal Diseases with Total Limbal Stem Cell Deficiency

Cited by 34 publications

References 24 publications

The growth-promoting effect of KGF on limbal epithelial cells is mediated by upregulation of ΔNp63α through the p38 pathway

The growth-promoting effect of KGF on limbal epithelial cells is mediated by upregulation of ΔNp63α through the p38 pathway

Human limbal epithelium contains side population cells expressing the ATP-binding cassette transporter ABCG2

Correlation of Cell Cycle Kinetics with p63 Expression in Human Limbal Epithelial Cells Expanded on Intact Human Amniotic Membrane

Contact Info

Product

Resources

About