EpiLife™ Medium, with 60 µM calcium

Mammalian cell culture media HEK

Experiment
Mammalian cell culture media HEK
Product
EpiLife™ Medium, with 60 µM calcium from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells, at 37°C and 10% CO2 in a 3:1 mixture of the DMEM and Ham’s F12 medium supplemented with 10% FCS, 5 µg/ml insulin, 0.4 µg/ml hydrocortisone, 10−10 M cholera toxin, and 2 × 10−9 M triiodothyronine

Publication protocol

Normal human epidermal keratinocytes were isolated from neonatal skin. Frozen keratinocytes were thawed and cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells, at 37°C and 10% CO2 in a 3:1 mixture of the DMEM and Ham’s F12 medium supplemented with 10% FCS, 5 µg/ml insulin, 0.4 µg/ml hydrocortisone, 10−10 M cholera toxin, and 2 × 10−9 M triiodothyronine, as described previously (Rheinwald and Green, 1975; Rochat et al., 1994). Human epidermal keratinocytes were also cultivated with CnT-PR medium (CELLnTEC), EpiLife medium containing supplement S7 (Life Technologies), and MCDB153 medium containing bovine pituitary extract (Hashimoto et al., 1994). Keratinocytes were used between passages 2 and 7. The medium was changed every 4 d. Cultures were fixed with 3.7% buffered formaldehyde and stained with 1% rhodamine B, and keratinocyte colonies were counted under a binocular microscope (SMZ645; Nikon). Clonal analysis was performed as described previously (Barrandon and Green, 1987b). In brief, human keratinocytes were cultivated at clonal density on a feeder layer of irradiated or mitomycin C–treated 3T3-J2 cells. Well-isolated colonies were then individually trypsinized in a cloning ring and subcultured into a 10-cm cell culture dish so clonal types could be evaluated.

Full paper   Login or join for free to view the full paper.

Reviews

EpiLife™ Medium, with 60 µM calcium from Thermo Fisher Scientific has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Mammalian cell culture media HEK using EpiLife™ Medium, with 60 µM calcium from Thermo Fisher Scientific.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for EpiLife™ Medium, with 60 µM calcium below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Mammalian cell culture media HEK using EpiLife™ Medium, with 60 µM calcium from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms