Cell culture factors influencing in vitro expression of mouse mammary tumor virus |
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Authors: | D L Fine L O Arthur L J T Young |
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Institution: | (1) RNA Virus Laboratory, NCI Frederick Cancer Research Center, Box B, 21701 Frederick, Maryland;(2) Department of Pathology, School of Medicine, University of California, 95616 Davis, California |
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Abstract: | Summary Several cell culture factors were found to influence in vitro expression of mouse mammary tumor virus (MMTV) in the mouse
adenocarcinoma cell line Mm5mt/c1. Cells were propagated in a variety of commercially available cell culture media to which dexamethasone (DXM) was added as
a stimulator of MMTV production. Culture seeding density, culture medium type, and glucose concentration each influenced MMTV
production when expressed on a per cell basis. Maximum cell growth occurred in cultures grown in RPMI-1640 medium containing
insulin. Those media which provided good cell growth were not necessarily optimal for virus expression. Addition of insulin
did not stimulate MMTV synthesis although dexamethasone alone was stimulatory in all media used; however, maximum MMTV expression
was obtained when dexamethasone and insulin were used in concert. Equivalent levels of MMTV-specific cell membrane antigen,
MMTV-specific protein, and virus particles were produced at incubation temperatures of 32°, 34° or 37° C; however, higher
levels of virus-related RNA-dependent DNA polymerase (RDDP) activity were recovered from cultures incubated at 32° and 34°
C than at 37° C. Decreased levels of RDDP were attributed to enzyme thermolability at 37° C incubation.
Research sponsored by the National Cancer Institute under Contract No. N01-CO-25423 with Litton Bionetics, Inc., and Contract
No. N01-CP-33253 with the University of California. |
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Keywords: | mouse mammary tumor virus cell culture dexamethasone virus expression hormonal stimulation RNA-dependent-DNA polymerase |
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