Two different human cell lines were employed to assess the in vitro tolerance of the developed preparations. Immortalized human corneal epithelial cells (HCECs; Evercyte GmbH, Vienna, Austria) were maintained at 37 °C under 5% CO2 in a humid atmosphere (95%). The medium was changed every 48–72 h. HCECs were kept in an EpiLife® cell-culture medium (Life Technologies, Madrid, Spain) with EDGS® 1X (Life Technologies, Madrid, Spain) and penicillin–streptomycin 1% (Life Technologies, Madrid, Spain) as supplementation. Likewise, immortalized human conjunctival epithelial cells (HConEpiCs; Innoprot, Bizkaia, Spain) were maintained under the same temperature, humidity, and CO2 conditions, while the medium was changed every 48 h. HConEpiCs were cultured using the IM-Ocular Epithelial Cell Medium Kit (Innoprot, Bizkaia, Spain). To ensure correct cell attachment and maintenance, the flasks were coated with collagen I (1 mg/mL) (Innoprot, Bizkaia, Spain).
Im ocular epithelial cell medium kit
Manufactured by Innoprot
Sourced in Spain
The IM-Ocular Epithelial Cell Medium Kit is a cell culture medium designed to support the growth and maintenance of ocular epithelial cells. The kit provides the necessary components to cultivate these specialized cells in an in vitro environment.
Automatically generated - may contain errors
2 protocols using im ocular epithelial cell medium kit
1
In Vitro Tolerance of Human Cell Lines
2
Isolation and Culture of Conjunctival Cells
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Primary conjunctival fibroblasts were obtained after digestion of VKC tarsal biopsies with 20-U/mL type I collagenase (Worthington Biochemical, Lakewood, NJ) at 37°C for 6 hours. Immortalized Human Conjunctival Epithelial (P10780-IM; Innoprot, Derio (Bizkaia), Spain) and the human monocytes U937 (Thermo Scientific; Wilmington, DE, USA) were purchased to be used for the experiments. Conjunctival epithelial cells were cultured using the IM-Ocular Epithelial Cell Medium Kit (Innoprot), and fibroblasts were grown in complete Dulbecco's modified Eagle's medium (DMEM), both supplemented with 10% fetal bovine serum, 1% (v/v) penicillin–streptomycin, and 2-mM glutamine at 37°C, in a 5% humidified CO2 atmosphere. The U937 cells were grown in suspension in complete Gibco RPMI 1640 Medium (Thermo Fisher Scientific) and were subcultured twice weekly by dilution using a seeding density of 106 cells.
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