Abstract
Limbal stem cells (LSC) have an important role in the maintenance of the corneal surface epithelium, and autologous cultured limbal epithelial cell (HLECs) transplantations have contributed substantially to the treatment of the visually disabling condition known as LSC deficiency. A major challenge is the ability to identify LSC in vitro and in situ, and one of the major controversies in the field relates to reliable LSC markers. This study was carried out to evaluate the culture of a limbal biopsy on human amniotic membrane (HAM): directly on the chorionic side and on intact epithelium, and the expression of the stem cell associated markers: ABCG2, p63. HAM has been extensively used for ocular surface reconstruction and has properties which facilitate the growth of epithelial cells controlling inflammation and scarring.
References
Baharvand H, Heidari M et al (2007) Proteomic analysis of epithelium-denuded human amniotic membrane as a limbal stem cell niche. Mol Vis 13:1711–1721
Chen Z, De Paiva C et al (2004) Characterization of putative stem cell phenotype in human timbal epithelia. Stem Cells 22:355–366
De Paiva CS, Chen Z et al (2005) ABCG2 transporter identifies a population of clonogenic human limbal epithelial cells. Stem Cells 23:63–73
Díaz-Prado S, Rendal-Vázquez ME et al (2010) Potential use of the human amniotic membrane as a scaffold in human articular cartilage repair. Cell Tissue Bank 11:183–195
Grueterich M, Tseng SC (2002) Human limbal progenitor cells expanded on intact amniotic membrane ex vivo. Arch Ophthalmol 120:783–790
Grueterich M, Espana EM et al (2003) Ex vivo expansion of limbal epithelial stem cells: amniotic membrane serving as a stem cell niche. Surv Ophthalmol 48:631–646
Koizumi N, Rigby H et al (2007) Comparison of intact and denuded amniotic membrane as a substrate for cell-suspension culture of human limbal epithelial cells. Graefes Arch Clin Exp Ophthalmol 245(1):123–134 [Epub 2006 Apr 13]
Kruse FE (1994) Stem cells and corneal epithelial regeneration. Eye 8:170–183
Li W, He H et al (2006) Basement membrane dissolution and reassembly by limbal corneal epithelial cells expanded on amniotic membrane. Invest Ophthalmol Vis Sci 47:2381–2389
Mariappan I, Maddileti S et al (2010) In vitro culture and expansion of human limbal epithelial cells. Nat Protoc 5:1470–1479
Nakamura T, Koizumi N et al (2003) Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in several ocular surface disease. Cornea 22:70–71
Nakamura T, Inatomi T et al (2006) Transplantation of autologous serum-derived cultivated corneal epithelial equivalents for the treatment of severe ocular surface disease. Ophthalmology 113:1765–1772
Parsa R, Yang A et al (1999) Association of p63 with proliferative potential in normal and neoplastic human keratinocytes. J Invest Dermatol 113:1099–1105
Pellegrini G, Dellambra E et al (2001) p63 identifies keratinocyte stem cells. Proc Natl Acad Sci USA 98:3156–3161
Schlotzer-Schrehardt U, Kruse FE (2005) Identification and characterization of limbal stem cells. Exp Eye Res 81:247–264
Tseng SC, Meller D et al (2002) Ex vivo preservation and expansion of human limbal epithelial stem cells on amniotic membrane for treating corneal diseases with total limbal stem cell deficiency. Adv Exp Med Biol 506:1323–1334
Zhang X, Sun H et al (2005) Comparison of cell-suspension and explant culture of rabbit limbal epithelial cells. Exp Eye Res 80:227–233
Zhou M, Li XM et al (2006) Transcriptional profiling of enriched populations of stem cells versus transient amplifying cells: a comparison of limbal and corneal epithelial basal cells. J Biol Chem 281:19600–19609
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Rendal-Vázquez, M.E., San-Luis-Verdes, A., Yebra-Pimentel-Vilar, M.T. et al. Culture of limbal stem cells on human amniotic membrane. Cell Tissue Bank 13, 513–519 (2012). https://doi.org/10.1007/s10561-012-9300-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10561-012-9300-x