Corneal epithelial cultures generated from organ-cultured limbal tissue: factors influencing epithelial cell growth.


PURPOSE:To explore the in vitro proliferative potential of human limbal epithelial cells after 31 degrees C organ-culture storage and to investigate putative factors influencing it. METHODS:185 cultures of limbal explants were carried-out either from full-thickness explants (n = 102) or from enzymatically dissociated cells (n = 83) seeded on a feeder layer of human keratocytes. Epithelial outgrowth was assessed by phase contrast microscopy using a computerized image analysis software. Cell phenotype was evaluated by transmission electron microscopy and immunocytology. Univariate and multivariate analysis were performed to determine factors influencing epithelial growth in culture. RESULTS:An epithelial outgrowth of 100 square mm or more was observed in 52% of cultures, (average growth area: 440 +/- 256 mm at three weeks). Corneal epithelial phenotype was confirmed by transmission electron microscopy, and cytokeratin pattern. Cytokeratine 19, deltaNp63, nestin and vimentin positive staining revealed undifferentiated epithelial cells in both explant and cell suspension cultures at three weeks. Short death to cornea retrieval time (p < 0.03) and female donors (p < 0.01) were associated with higher cell growth. Enzymatic treatment of explants by trypsin, but not dispase, decreased cell proliferation at two (p < 0.03) and three weeks (p < 0.04). Donor age, duration of corneal storage, and source of the explant did not influence the cell growth. CONCLUSION:Organ-culture conditions can preserve limbal cell mitotic potential if limbal tissue is excised early after circulatory arrest. Human keratocytes can be used as a feeder layer allowing epithelial cells to maintain poorly differentiated phenotype in culture. Further investigations are needed to explain the influence of the donor sex on epithelial cell growth in culture.


Curr Eye Res


Current eye research


Zito-Abbad E,Borderie VM,Baudrimont M,Bourcier T,Laroche L,Chapel C,Uzel JL




Has Abstract


2006-05-01 00:00:00














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    journal_title:Current eye research

    pub_type: 杂志文章


    authors: Gella L,Raman R,Sharma T

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  • A comparison of three methods for trephining donor corneal buttons: endothelial cell loss and microscopic ultrastructural evaluation.

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    authors: Moshirfar M,Meyer JJ,Kang PC

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  • Long-Term Changes in Straylight Induced by Overnight Orthokeratology: An Objective Measure Using the Double-Pass System.

    abstract::Purpose: To map the time course of changes in intraocular straylight of the human eye 1 year after initial lens wearing for orthokeratology treatment using the objective double-pass technique. Materials and Methods: A total of 35 subjects (19 males and 16 females) completed the study. The mean age was 11.46 ± 2.33 yea...

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    authors: Liu G,Jin N,Bi H,Du B,Gu T,Zhang B,Wei R

    更新日期:2019-01-01 00:00:00

  • In Vivo Efficacy of Histatin-1 in a Rabbit Animal Model.

    abstract::Purpose/Aim: Corneal abrasions and nonhealing corneal epithelial defects are common conditions that cause pain and sometimes are slow to heal. Histatins, a family of histidine-rich peptides, have been implicated in oral and skin epithelial wound healing, and have been shown to be effective in vitro in human corneal ep...

    journal_title:Current eye research

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    authors: Oydanich M,Epstein SP,Gadaria-Rathod N,Guers JJ,Fernandez KB,Asbell PA

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  • Aqueous humor levels of topically applied levofloxacin in human eyes.

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    authors: Yamada M,Mochizuki H,Yamada K,Kawai M,Mashima Y

    更新日期:2002-05-01 00:00:00

  • The quantity of rhodopsin in human eyes.

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    journal_title:Current eye research

    pub_type: 杂志文章


    authors: Fulton AB,Dodge J,Schremser JL,Armstrong A,Lanier F,Dawson WW,Williams TP

    更新日期:1990-12-01 00:00:00

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    journal_title:Current eye research

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    pub_type: 临床试验,杂志文章


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    authors: Li LK,Roy D,Spector A

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    journal_title:Current eye research

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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    journal_title:Current eye research

    pub_type: 杂志文章


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    journal_title:Current eye research

    pub_type: 杂志文章


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    journal_title:Current eye research

    pub_type: 杂志文章


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    journal_title:Current eye research

    pub_type: 杂志文章


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    journal_title:Current eye research

    pub_type: 杂志文章


    authors: Lipman RD,Muggleton-Harris AL

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