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.
Age
embryo
Applications
Mitomycin C-treated cells are used as a feeder layer
Storage Conditions
liquid nitrogen vapor phase
Comments
These cells are generated from mouse embryonic fibroblasts, STO cell line (ATCC CRL-1503), by a treatment of Mitomycin C.
These cells are provided to be used as feeder cells to support the growth of other cells. They have been treated with Mitomycin C and will not replicate. The cells will begin to deteriorate in 2 to 3 weeks after plating. Once the feeder cells have attached, the culture medium can be changed to accommodate the cells to be supported. Such populations are employed for maintenance of embryonal stem cells such as ES-D3 (ATCC CRL-1934) or teratocarcinoma stem cells (see ATCC CRL-1535 and ATCC CRL-1566) in the undifferentiated state.
It is recommended that the feeder cells be plated 24 hours before use at 6 x 106/T-75 or 2 x 106/T-25 in order to obtain a 100% confluent monolayer for stem cells growth.
Feeder layers are also employed to enhance the growth at low density populations of many hybridomas, colon carcinoma cell line (ATCC CCL-247) and squamous cell carcinoma cell lines (ATCC CRL-1624 and ATCC CRL-1629). This action is due partly to conditioning of the substrate and the medium. A feeder layer confluence of 30 % is adequate and can be obtained by plating 2 x 106/T-75.
Complete Growth Medium
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Cryopreservation
Freeze Medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
Name of Depositor
ATCC
References
Martin GR, Evans MJ. Differentiation of clonal lines of teratocarcinoma cells: formation of embryoid bodies in vitro. Proc. Natl. Acad. Sci. USA 72: 1441-1445, 1975. PubMed: 1055416
Teratomas and Differentiation, eds. Michael I. Sherman and David Solter, 13–14. New York: Academic Press, 1975.
Martin GR, et al. The development of cystic embryoid bodies in vitro from clonal teratocarcinoma stem cells. Dev. Biol. 61: 230-244, 1977. PubMed: 590624
