ISSN:
1573-0603
Keywords:
Differentiation
;
Mouse mammary gland
;
Retroviral vector
;
Stem cell
;
Whole organ culture
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract The objective of this study was to develop a method to indelibly mark mammary cells to follow cell lineage. This was done by infecting mouse mammary glands in whole organ culture (WOC) with a retroviral vector containing the β-galactosidase (BAG) gene. Abdominal mammary glands from BALB/c mice primed with estrogen and progesterone were removed and injected with 30 µl of concentrated vector prepared from CRE BAG 2 (ATCC 1858 — CRL) cells or left uninjected and co-cultured with CRE BAG 2 feeder layers for 48 or 96 hours. Glands were floated on siliconized lens paper and incubated in Waymouth's 752/1 medium supplemented with insulin, aldosterone, hydrocortisone, prolactin, and epidermal growth factor for 5 days. Integration of BAG was determined with a colorimetric assay using X-gal, a β-galactoside analog, in whole mounts and collagenase digested glands. Blue cells in collagenase digests and blue areas in whole mounts indicated that BAG had integrated into mammary cells during whole organ culture.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00986230
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