Research-grade corneoscleral rims of human cadaver donors were procured from the eye bank CorneaGen Inc. (Seattle, WA, USA) and were handled in accordance with the tenets of the Declaration of Helsinki. Corneal limbal cells were carefully collected as previously described [9] (link). Cells were seeded at 1 × 104 cells/cm2 onto plastic tissue culture plates coated with 0.5 μg/cm2 laminin 511 (iMatrix-511, Nippi, Tokyo, Japan). Cells were cultured in corneal epithelium maintenance medium (CEM), as it is reported to be suitable for HCEC maintenance [20] , and incubated with 5% CO2 at 37 °C. CEM consists of DMEM:F-12 medium (1:1) containing l -glutamine and 2.438 g/L sodium bicarbonate (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 2% B-27 (Thermo Fisher Scientific), 10 μM of Y-27632 (Wako Pure Chemical Industries, Osaka, Japan), 10–20 ng/mL of human recombinant KGF/FGF-7 (R&D Systems, Minneapolis, MN, USA), and 1% Penicillin-Streptomycin (Thermo Fisher Scientific).
Human recombinant kgf fgf 7
Manufactured by R&D Systems
Sourced in Japan
Human recombinant KGF/FGF-7 is a growth factor protein produced in a recombinant expression system. It is a member of the fibroblast growth factor family and plays a role in cell proliferation and differentiation.
Automatically generated - may contain errors
Lab products found in correlation
Sodium bicarbonate, by Thermo Fisher Scientific (3 mentions)
Y 27632, by Fujifilm (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Imatrix 511, by Nippi (1 mentions)
Cacl2, by Fujifilm (1 mentions)
B27 supplement, by Thermo Fisher Scientific (1 mentions)
2 protocols using human recombinant kgf fgf 7
1
Culturing Human Corneal Epithelial Cells
2
Culturing Human Corneal Epithelial Cells
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Human corneal epithelial cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM): F12 Medium (1:1) (Life Technologies, Carlsbad, CA, USA) supplemented with a B-27® supplement (Life Technologies)` 10 μM of Y-27632 (Wako Pure Chemical Industries, Osaka, Japan), 20 ng/ml of human recombinant KGF/FGF-7 (R & D Systems, Minneapolis, MN, USA) and 2 mM of L-Glutamine (Life Technologies)42 (link), and incubated in 5% CO2 at 37 °C. Immortalized human keratinocytes (OKF6/TERT-1, OKF6/TERT-2, N/TERT-1, N/TERT-2 cells) were obtained from the laboratory of Dr. J. Rheinwald (Harvard Institutes of Medicine, Boston, MA, USA)19 (link). Cells were cultured in Keratinocyte-SFM supplemented with 25 μg/ml of bovine pituitary extract (BPE), 0.2 ng/ml of epidermal growth factors (EGF, Life Technologies) and 0.3 mM of CaCl2 (Wako Pure Chemical Industries). Again, the cells were incubated in 5% CO2 at 37 °C. The other human cells, which were used in the current study and their culture media are listed in Supplementary Table S6 .
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