Enhanced growth medium and method for culturing human mammary epithelial cells

Title: Enhanced growth medium and method for culturing human mammary epithelial cells

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Abstract

Methods are disclosed for isolating and culturing human mammary epithelial cells of both normal and malignant origin. Tissue samples are digested with a mixture including the enzymes collagenase and hyaluronidase to produce clumps of cells substantially free from stroma and other undesired cellular material. Growing the clumps of cells in mass culture in an enriched medium containing particular growth factors allows for active cell proliferation and subculture. Clonal culture having plating efficiencies of up to 40% or greater may be obtained using individual cells derived from the mass culture by plating the cells on appropriate substrates in the enriched media. The clonal growth of cells so obtained is suitable for a quantitative assessment of the cytotoxicity of particular treatment. An exemplary assay for assessing the cytotoxicity of the drug adriamycin is presented.

Inventors:

Stampfer, Martha R ^[1]; Smith, Helene S ^[2]; Hackett, Adeline J ^[3]

7290 Sayre Dr., Oakland, CA 94611
5693 Cabot Dr., Oakland, CA 94611
82 Evergreen Dr., Orinda, CA 94563

Issue Date:: 1983-01-01

Research Org.:: Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

OSTI Identifier:: 864822

Patent Number(s):: 4423145

Assignee:: Stampfer, Martha R. (7290 Sayre Dr., Oakland, CA 94611);Smith, Helene S. (5693 Cabot Dr., Oakland, CA 94611);Hackett, Adeline J. (82 Evergreen Dr., Orinda, CA 94563)

Patent Classifications (CPCs):: G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES

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G - PHYSICS G01 - MEASURING G01N - INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
G01N33/5014 - {for testing toxicity}

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N2502/23 - Gastro-intestinal tract cells

Y - NEW / CROSS SECTIONAL TECHNOLOGIES Y10 - TECHNICAL SUBJECTS COVERED BY FORMER USPC Y10S - TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
Y10S435/948 - using viruses or cell lines

C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
C12N2501/33 - Insulin
C12N2509/00 - Methods for the dissociation of cells, e.g. specific use of enzymes
C12N5/0631 - {Mammary cells}
C12N2502/253 - Bladder cells
C12N2501/01 - Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
C12N2503/02 - Drug screening

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Resource Type:: Patent

Country of Publication:: United States

Language:: English

Subject:: enhanced; growth; medium; method; culturing; human; mammary; epithelial; cells; methods; disclosed; isolating; normal; malignant; origin; tissue; samples; digested; mixture; including; enzymes; collagenase; hyaluronidase; produce; clumps; substantially; free; stroma; undesired; cellular; material; growing; mass; culture; enriched; containing; particular; factors; allows; active; cell; proliferation; subculture; clonal; plating; efficiencies; 40; obtained; individual; derived; appropriate; substrates; media; suitable; quantitative; assessment; cytotoxicity; treatment; exemplary; assay; assessing; drug; adriamycin; cellular material; tissue sample; individual cells; substantially free; individual cell; medium containing; mixture including; tissue samples; growth medium; epithelial cells; human mammary; mammary epithelial; active cell; culturing human; /435/

Citation Formats


                    Stampfer, Martha R, Smith, Helene S, and Hackett, Adeline J. Enhanced growth medium and method for culturing human mammary epithelial cells.  United States: N. p., 1983. 
        Web.

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                    Stampfer, Martha R, Smith, Helene S, & Hackett, Adeline J. Enhanced growth medium and method for culturing human mammary epithelial cells.  United States.

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                    Stampfer, Martha R, Smith, Helene S, and Hackett, Adeline J. Sat .  
        "Enhanced growth medium and method for culturing human mammary epithelial cells".  United States.  https://www.osti.gov/servlets/purl/864822.

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@article{osti_864822,

  title        = {Enhanced growth medium and method for culturing human mammary epithelial cells},

  author       = {Stampfer, Martha R and Smith, Helene S and Hackett, Adeline J},

  abstractNote = {Methods are disclosed for isolating and culturing human mammary epithelial cells of both normal and malignant origin. Tissue samples are digested with a mixture including the enzymes collagenase and hyaluronidase to produce clumps of cells substantially free from stroma and other undesired cellular material. Growing the clumps of cells in mass culture in an enriched medium containing particular growth factors allows for active cell proliferation and subculture. Clonal culture having plating efficiencies of up to 40% or greater may be obtained using individual cells derived from the mass culture by plating the cells on appropriate substrates in the enriched media. The clonal growth of cells so obtained is suitable for a quantitative assessment of the cytotoxicity of particular treatment. An exemplary assay for assessing the cytotoxicity of the drug adriamycin is presented.},

  doi          = {},

  journal      = {},
number       = ,

  volume       = ,

  place        = {United States},

  year         = {1983},

  month        = {1}

}

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