Limbal Explants from Cryopreserved Cadaver Human Corneas. Immunofluorescence and Light Microscopy of Epithelial Cells Growing in Culture

Authors: Bratanov, M.; Neronov, A.; Nikolova, E.

Source: Cryoletters, Volume 30, Number 3, May 2009 , pp. 183-189(7)

Buy & download fulltext article:

Price: $26.98 plus tax (Refund Policy)

Abstract:

The aim of the present study was to determine whether human cadaver corneas, that were subject to cryopreservation, would be a source of migrating epithelial cells in vitro and what kind of morphological features these cells possess. Limbal explant culture was used for expanding the epithelial cells. Non-quantitative light microscopical examinations of the cultures within a period of 28 days were carried out. The phenotype of cultured cells, particularly of the presumed adult stem cell population, was examined by indirect fluorescent immunostaining using antibodies against corneal stem cell associated markers p63 and vimentin. The effectiveness of the freezing-thawing protocol was confirmed by cultivation of limbal explants taken from non-cryopreserved cadaver corneoscleral rims.

The result clearly showed that limbal tissue, subjected to cryopreservation and long lasting (up to 12 months) storage in liquid nitrogen, retains the capacity to be source of migrating and proliferating epithelial cells in vitro including the presumed adult stem cells and transient amplifying cells.

Keywords: CRYOPRESERVATION; LIMBAL EXPLANT CULTURE; LIGHT MICROSCOPY; IMMUNOFLUORESCENCE; P63; VIMENTIN

Document Type: Research article

Publication date: 2009-05-01

More about this publication?

CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.

Limbal Explants from Cryopreserved Cadaver Human Corneas. Immunofluorescence and Light Microscopy of Epithelial Cells Growing in Culture

Buy & download fulltext article:

Tools

Key

Text size:

Useful pages