Enhancement of differentiation of human lens epithelium in tissue culture by changes in cell-substrate adhesion

• Published in: Investigative ophthalmology & visual science Vol. 31; no. 11; pp. 2395 - 2404
• Main Authors: Arita, T, Lin, LR, Susan, SR, Reddy, VN
• Format: Journal Article
• Language: English
• Published: Rockville, MD ARVO 01.11.1990; Association for Research in Vision and Ophtalmology
• Subjects: adhesion; Adult; Aged; Aquaporins; Biological and medical sciences; Cell Adhesion - physiology; Cell Differentiation; cells; Crystallins - biosynthesis; Culture Techniques - methods; Epithelial Cells; epithelium; Epithelium - ultrastructure; eye; Eye and associated structures. Visual pathways and centers. Vision; eye lens; Eye Proteins - biosynthesis; Fundamental and applied biological sciences. Psychology; gamma -crystallin; gap junctions; Gene Expression; Humans; Intercellular Junctions - ultrastructure; Lens, Crystalline - cytology; Lens, Crystalline - ultrastructure; man; Membrane Glycoproteins; Middle Aged; Reproducibility of Results; substratum; tissue culture; Vertebrates: nervous system and sense organs; Human; Cell culture; Scanning electron microscopy; Cell cell interaction; Cell differentiation; Gene expression; In vitro; Eye; Visual system; Transmission electron microscopy; Ultrastructure; Lentoid body; Epithelial cell; Lens
• ISSN: 0146-0404; 1552-5783

Differentiation of human lens epithelial (HLE) cells cultured in vitro was drastically accelerated when the cells were cultured on cell-substrate adhesion-free surfaces. Spontaneous differentiation of the lens epithelial cells in monolayer cultures could be recognized with the appearance of lentoid bodies after 40-50 days if maintained without further passage. Although dissociated HLE cells reconstituted into monolayers consistently on the haptotactic substrates (either gold-coated biopore membrane or regular plastic dishes), the cells from the same batches exclusively formed cell aggregates when cultured on either biopore membrane or agarose-coated plastic dishes (nonhaptotactic). The cells on nonadhesion substrate first aggregate, then synchronously develop into lentoids by the 10th day of culture. The differentiation of HLE cells into lentoid structures with lens-fiber characteristics was documented by both ultrastructural and biochemical markers, such as loss of cytoplasmic organelles, formation of gap junctions, and the expression of gamma-crystallin and MP26. The system, in which differentiation of epithelial cells can be induced predictably in a short period of time, provides an excellent model for the study of differentiation and gene expression in human lens cells cultured in vitro.