|Catalog. No.||Product Name||Format||Stock Conc.||Storage on Arrival||Thawing Instructions||Storage Once Thawed|
|ax336894||Primary Human Epithelial Colon||500,000 cells/vial||N/A||Place into vapor||Follow||N/A|
|Cells – Ascending Colon||phase liquid nitrogen storage||Protocol|
|ax3368941||Primary Human Epithelial Colon||500,000 cells/vial||N/A||Place into vapor||N/A|
|Cells – Descending Colon||phase liquid nitrogen storage||FollowProtocol|
|ax3368942||Primary Human Epithelial Colon||500,000 cells/vial||N/A||Place into vapor||Follow||N/A|
|Cells – Duodenum||phase liquid nitrogen storage||Protocol|
|ax3368943||Primary Human Epithelial Colon||500,000 cells/vial||N/A||Place into vapor|
|Cells – Transverse Colon||phase liquid nitrogen storage||FollowProtocol||N/A|
|ax336897||Human Intestine Cell Media||100mL||N/A||Stored at -20°C until ready to use||Media shouldbe warmed at||Stored at +4°C and used within 30 days|
|37°C prior to|
|use with cells|
- For intestinal epithelial cells to effectively attach and spread on tissue culture dishes, coating the bottom of the plates with Type I Collagen. Make sure the bottom of each well is completely covered.
- The recommended seeding density for intestinal epithelial cells is dependent on the colon region:
- Ascending Colon: ~25,000 cells/cm2
- Descending Colon: ~60,000 cells/cm2
- Duodenum: ~60,000 cells/cm2Transverse Colon: ~60,000 cells/cm
Thawing and plating of Epithelial Colon Cells
- Coat the culture plates with Type I Collagen or use pre-coated culture plates.
- Place 5mL Axol’s Intestine Cell Medium into 15mL centrifuge tube.
- Rapidly thaw the vial of frozen cells in a 37°C water bath until just prior to complete thawing (slurry of residual ice should be present). Thawing for longer than 2 minutes compromises cell viability. Wipe the outside of the vial with 70% ethanol.
- Transfer the cell suspension into the 15mL centrifuge tube containing 5mL of Intestine Cell Medium. Rinse the cryovial with 1mL Intestine Cell Medium and add it to the 15mL centrifuge tube.
- Centrifuge the tube at 600xg for 1 minute at room temperature to pellet the cells.
- Carefully aspirate the supernatant and resuspend in the volume of Intestine Cell Medium for the desired cell concentration and number of wells.
- Plate the desired number of cells in each well by adding the media directly to the middle of each well, being careful to not touch the bottom of each well.
Culture of Epithelial Colon Cells
- After plating, incubate cells in 5% CO2, 37°C tissue culture incubator overnight.
- Change media 24 hours after initial plating using pre-warmed Axol’s Intestine CellMedium and every 48 hours after that.
- Continue incubation at 37°C.
- Cells will be >50% confluency on day 5-7 after plating
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