#76 KhGH (CRL-8858)

Organism: Homo sapiens, human  /  Cell Type: keratinocyte  / 

Permits and Restrictions

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Organism Homo sapiens, human
Cell Type keratinocyte
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 CELLS CONTAIN RETROVIRUS, RECOMBINANT

Biosafety  classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Applications
The cells produce and secrete hGH.
Recovery of the cells from the frozen state, and subsequent proliferation, may be enhanced by the use of a feeder layer of irradiate or mitomycin C treated 3T3 cells.
IRR-3T3 (ATCC 48-X) may be used as a feeder layer for this cell line.
Genes Expressed
human growth hormone (hGH)
Cellular Products
human growth hormone (hGH)
Tumorigenic No
Effects
No, The cells were not tumorigenic in immunosuppressed mice, but did form colonies in semisolid medium.
Comments
The cells contain a recombinant retrovirus genome that contains sequences coding for hGH and neomycin resistance.
The cells produce and secrete hGH.
Recovery of the cells from the frozen state, and subsequent proliferation, may be enhanced by the use of a feeder layer of irradiate or mitomycin C treated 3T3 cells.
IRR-3T3 (ATCC 48-X) may be used as a feeder layer for this cell line.
Complete Growth Medium A 3:1 mixture of Ham's F12 (hypoxanthine free) medium and Dulbecco's modified Eagle's medium supplemented with 400 ng/ml hydrocortisone, 0.01 mg/ml cholera toxin, 0.005 mg/ml insulin and 10 ng/ml epidermal growth factor, 95%; fetal bovine serum, 5%
Subculturing
Protocol: Remove spent medium, rinse cell with fresh 0.25% trypsin, remove trypsin and allow the culture to stand for 5 to 10 minutes at 37C. Add fresh medium, aspirate and dispense into new flasks.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Name of Depositor Whitehead Institute for Biomedical Research
U.S. Patent Number
Disclosure This material is cited in a US or other Patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Patent Depositor of the party to which the material was furnished.
References

Morgan JR, Mulligan RC. Introduction and expression of foreign genetic material in epithelial cells. US Patent 4,868,116 dated Sep 19 1989

Morgan JR, Mulligan RC. In vivo introduction and expression of foreign genetic material in epithelial cells. US Patent 4,980,286 dated Dec 25 1990

Allen-Hoffmann BL, Rheinwald JG. Polycyclic aromatic hydrocarbon mutagenesis of human epidermal keratinocytes in culture. Proc. Natl. Acad. Sci. USA 81: 7802-7806, 1984. PubMed: 6440145

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Morgan JR, Mulligan RC. Introduction and expression of foreign genetic material in epithelial cells. US Patent 4,868,116 dated Sep 19 1989

Morgan JR, Mulligan RC. In vivo introduction and expression of foreign genetic material in epithelial cells. US Patent 4,980,286 dated Dec 25 1990

Allen-Hoffmann BL, Rheinwald JG. Polycyclic aromatic hydrocarbon mutagenesis of human epidermal keratinocytes in culture. Proc. Natl. Acad. Sci. USA 81: 7802-7806, 1984. PubMed: 6440145