Cell culture quality control by rapid isoenzymatic characterization

Halton, David M.; Peterson, Ward D.; Hukku, Bharati

doi:10.1007/bf02617989

Cited by 44 publications

(23 citation statements)

References 20 publications

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“…Samples were applied to the slots of the agarose film. The films were placed in an electrophoresis chamber and run for 25 min and stained by using colorimetric reactions according to a protocol presented elsewhere (19). The cell extract from a cell line of mouse origin was used as a negative control.…”

Section: Vol 76 2002mentioning

confidence: 99%

Development and Characterization of Bovine × Human Hybrid Cell Lines That Efficiently Support the Replication of both Wild-Type Bovine and Human Adenoviruses and Those with E1 Deleted

Olphen

Mittal

2002

J Virol

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The 293 cell line that was generated by transforming human embryonic kidney cells with human adenovirus type 5 (HAV5) early region 1 (E1) sequences is an excellent host for generating and growing HAV5 recombinants with E1 deleted, but it does not support the replication of bovine adenovirus type 3 (BAV3). Madin-Darby bovine kidney (MDBK), an established bovine cell line, is an excellent host for growing and plaquing BAV3. For the purpose of combining the unique characteristics of these two cell lines (293 and MDBK), we generated a number of bovine ؋ human hybrid (BHH) cell lines. Comparison of three BHH hybrid clones-BHH3, BHH8, and BHH2C-with 293-Puro (puromycin-resistant 293 cells) and MDBK-Neo (G418-resistant MDBK cells) cell lines for total cellular DNA content, species-specific surface markers, isoenzyme analysis, and karyotyping indicate that they are hybrid in nature. BHH clones constitutively expressed the E1 proteins (E1A, E1B-21kDa, and E1B-55kDa) of HAV5 and efficiently supported the replication of both wild-type and replication-incompetent bovine or human adenoviruses. Transient gene expression experiments with a plasmid encoding the bacterial ␤-galactosidase gene demonstrated that BHH cell hybrids seem to have better transfection efficiencies than either of the parental cell lines. These cell lines will be useful for isolating and growing replication-competent human or bovine adenovirus recombinants with E1 deleted and for the study of cellular or viral factors important for viral replication. The development of somatic cell hybrids appears to be a simple way of combining some of the desirable characteristics present separately in two parental cell lines.The most widespread use of somatic cell hybrids is the formation of hybridomas for monoclonal antibody production. They are generated by the fusion of antibody-producing lymphocytes from a mouse or rat immunized with an antigen of interest with a myeloma cell line (1, 29). Production of interand intraspecific cell hybrids by means of polyethylene glycol (PEG)-induced fusion for extending the life of mammalian cells has been demonstrated (42). Somatic cell hybrids have also been used as tools for studying the chromosomal assignment of genes (12,26,27), mapping normal as well as tumorrelated genes (47), and localizing the site-specific integration of adeno-associated proviral DNA into human chromosome 19 (31, 49). Hybridization between bovine ϫ rodent, mouse, or hamster were exploited for the mapping of bovine genes (51), and bovine ϫ mouse hybrid cells were used for studying the replication of mengo virus (7,30,55).Adenoviral early region 1 (E1) genes are the first set of the genes that are expressed and are necessary for virus replication (22, 50). Adenovirus recombinants with E1 deleted are routinely produced and grown in a permissive cell line that constitutively expresses E1 proteins (17). The 293 cell line constitutively expresses E1 proteins and was derived from human embryonic kidney cells after transformation with human adenovirus type 5 (HA...

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Section: Vol 76 2002mentioning

confidence: 99%

Development and Characterization of Bovine × Human Hybrid Cell Lines That Efficiently Support the Replication of both Wild-Type Bovine and Human Adenoviruses and Those with E1 Deleted

Olphen

Mittal

2002

J Virol

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“…184, 184A1 cells (43rd passage), 184B5 cells (18th passage), and untreated 184 cells (7th passage) were analyzed for their profile of seven different polymorphic isozymes (23). All three cell types had an identical profile; the probability that such a result would occur by chance is <0.01%.…”

mentioning

confidence: 99%

Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzo[a]pyrene.

Stampfer¹,

Bartley²

1985

Proc. Natl. Acad. Sci. U.S.A.

347

236

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Rapidly growing primary cultures of normal human mammary epithelial cells (HMEC) were exposed to 1 jig of benzo [a]pyrene (B[a]P) per ml for two or three 24-hr periods. The B[a]P-treated populations consistently contained cells displaying a longer period of active growth in culture compared to the untreated control cells. Widespread heterogeneity in morphology and growth patterns was evidenced in these "extended life" (EL) cultures, with multiple sequential changes in these parameters occurring during the course of their life in culture. Two apparently immortal continuous cell lines have thus far emerged from these EL cultures. These lines have been characterized to be of human mammary epithelial origin and derived from the originally treated HMEC specimen. The continuous lines do not appear to be malignantly transformed as they do not cause tumor formation in nude mice and show little or no anchorage-independent growth. Nonetheless, they have acquired several properties characteristic of tumor-derived HMEC, which distinguish them from their normal progenitors. These cell lines, as well as the EL strains, may provide useful substrates for studies to determine what agents can induce further transforming events. Additionally, analysis of the multiple steps occurring in the EL cultures, as well as in the emergence of the continuous cell lines, could potentially elucidate the processes occurring during human epithelial cell carcinogenesis and escape from senescence. Establishment of methods

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“…It is considerably faster t h a n t h a t we previously described (13,14) and also provides a high degree of reproducibility. The method requires the use of control phenotype enzyme extracts prepared from appropriately characterized cell lines that are run in wells adjacent to the unknown cell extract.…”

Section: Discussionmentioning

confidence: 92%

“…system for isozymic phenotype analysis (13,14). We report here an even simpler and quicker derivative method for determining the phenotypes of G6PD, PGM1, PGM3, ESTD, Me-2, AK-1, and GLO-1 in human cell lines.…”

Section: Introductionmentioning

confidence: 99%

Rapid isoenzyme analysis of cell cultures by agarose electrophoresis II. Intraspecies identification of human cell lines

Ottenbreit

Halton

PetersonJr.

1983

Journal of Tissue Culture Methods

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Cell culture quality control by rapid isoenzymatic characterization

Cited by 44 publications

References 20 publications

Development and Characterization of Bovine × Human Hybrid Cell Lines That Efficiently Support the Replication of both Wild-Type Bovine and Human Adenoviruses and Those with E1 Deleted

Development and Characterization of Bovine × Human Hybrid Cell Lines That Efficiently Support the Replication of both Wild-Type Bovine and Human Adenoviruses and Those with E1 Deleted

Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzo[a]pyrene.

Rapid isoenzyme analysis of cell cultures by agarose electrophoresis II. Intraspecies identification of human cell lines

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