Cell line/type | Corneal epithelial cells |
---|---|
Species | Human |
Animal free | No |
Product | DMEM-F12 Yokoo et al. 2008 |
Yokoo, S., Yamagami, S., Usui, T., Amano, S., & Araie, M. (2008). Human corneal epithelial equivalents for ocular surface reconstruction in a complete serum-free culture system without unknown factors. Investigative ophthalmology & visual science, 49(6), 2438-2443. The purpose of this study was to establish a culture technique for human corneal epithelial equivalents that do not require fetal bovine serum (FBS), feeder cells, or bovine pituitary extracts and compare this system with conventional culture medium with FBS and mouse 3T3 fibroblasts. Human corneal limbal tissue from donor corneas was dissociated on denuded amniotic membranes and then cultured for 3 weeks in feeder-cell- and serum-free medium containing epidermal growth factor and B-27. Then, the cell sheet was evaluated by light microscopy, immunohistochemistry, and electron microscopy. The epithelial proliferative capacity was compared between serum- and feeder-cell-free medium and conventional medium. A stratified cell sheet expressing cytokeratin-3 and -12 was grown in serum- and feeder-cell-free medium without unknown growth factors. The epithelial proliferative capacity in feeder-cell- and serum-free medium determined by WST-1 and colony-forming efficiency was significantly higher than that in conventional medium. This method using serum- and feeder-cell-free medium not containing unknown growth factors allows the highly proliferative culture of human corneal epithelium. Serum-, feeder cell-, and BPE-free culture medium containing B-27 and EGF made it possible to grow human corneal limbal epithelial cell equivalents. This method allows the highly proliferative culture of HCE in serum- and feedercell– free medium, in comparison with conventional medium containing FBS cocultured with mouse 3T3 fibroblast feeder cells. The formulation of the serum-free, DMEM-F12 based medium is listed in the in the Materials and Methods, under the heading Culture of Human Corneal Limbal Epithelium. See Table 1. For the composition of B27 Medium Supplement. https://iovs.arvojournals.org/data/journals/iovs/932949/z7g00608002438.pdf |
|
Source | Literature - own formulation |
Chemically defined > Yes | Yes |
Contains phenol red > Yes | Yes |
Antibiotics free > No | No |