PHK16-0b Cell Line永生化人皮肤角化细胞株-Biovector NTCC典型培养物保藏中心

PHK16-0b Cell Line

Cat No.: NTCC925769

Description: NTCC® PHK16-0bcell line is a normal keratinocyte cellline immortalized with HPV16 virus genome and the growth is dependent to EGF. More than 95% of cells are diploid.

Species: human

Tissue: skin

Growth Properties:

Morphology: keratinocyte

Growth Medium: MCDB153 medium (0.03 mM Ca2+) with 5 ug/ml insulin, 0.5 ug/ml hydrocortisone, 10 ug/ml transferrin, 0.1 mM phosphorylethanolamine, 0.1 mM ethanolamine, 10 ng/ml EGF and 40-70 ug/ml BPE. MCDB153 with < 0.5% FCS was also used in original report.

Subculturing Procedure: Cells are harvested with 0.1% trypsin and 0.01% EDTA. Medium change every 2 days and subculture every 10 days.

Images:

After you receive cell lines

Do not immerse in liquid nitrogen directly!!!

If cryotube or glass ampoule has been stored directly in liquid nitrogen (liquid phase), there is a risk of exploding imperfectly sealed cryotube or glass. It is recommended to preserve them in vapor phase of liquid nitrogen tank.

Thawing

Cell lines should be thawed rapidly.

Take out the ampoule from the styrene foam package and thaw the content within 2 min by shaking in warm (not higher than 37°C.) water. Use safety gloves and a face shield to avoid injury from possible explosion of the ampoule. It is safe to thaw them one by one, even if it takes longer.

Seed at 25 cm2 flask or 6 cm dish.

All operations should be performed under aseptic conditions. Sterilize the entire surface of the ampoule with gauze moist with 70% ethanol or cationic detergent sterilizer, and cut the neck off while wrapped in sterilized gauze. Although every ampoule has a groove around the neck for easy cutting off, a precaution may be required to open the ampoule because of its hard structure. Transfer the cell suspension to a centrifuge tube, add 10 ml of the medium specified in the catalog, centrifuge the mixture at 1000 rpm for 3 min, and discard the supernatant. Without washing, resuspend the cells in the same medium and culture them by the standard procedure. Unless otherwise specified, the appropriate culture volume would be 5 ml per ampoule in a 25 cm2 flask or 6 cm dish for tissue cultures. The cell density given in the attached document or in the catalog may differ from the actual density on thawing. Make sure of cell proliferation before proceeding to passage, because excessive dilution may lead to cell death particularly in hemocytes and lymphocytes.

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