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Vol. 16, No. 6, pp. 693-706, March 15, 2002
1 Structural Cell Biology Unit, Institute of Medical
Anatomy, The Panum Institute, DK-2200 Copenhagen N, Denmark;
2 Zoophysiological Laboratory, The August Krogh Institute,
DK-2100 Copenhagen Ø, Denmark; 3 Life Sciences Division,
Lawrence Berkeley National Laboratory, University of California,
Berkeley, California 94720, USA
The epithelial compartment of the human breast comprises two
distinct lineages: the luminal epithelial and the myoepithelial lineage. We have shown previously that a subset of the luminal epithelial cells could convert to myoepithelial cells in culture signifying the possible existence of a progenitor cell. We therefore set out to identify and isolate the putative precursor in the luminal
epithelial compartment. Using cell surface markers and immunomagnetic
sorting, we isolated two luminal epithelial cell populations from
primary cultures of reduction mammoplasties. The major population
coexpresses sialomucin (MUC+) and epithelial-specific antigen
(ESA+) whereas the minor population has a suprabasal position
and expresses epithelial specific antigen but no sialomucin
(MUC
/ESA+). Two cell lines were further
established by transduction of the E6/E7 genes from human papilloma
virus type 16. Both cell lines maintained a luminal epithelial
phenotype as evidenced by expression of the tight junction proteins,
claudin-1 and occludin, and by generation of a high transepithelial
electrical resistance on semipermeable filters. Whereas in clonal
cultures, the MUC+/ESA+ epithelial cell line was
luminal epithelial restricted in its differentiation repertoire, the
suprabasal-derived MUC/ESA+ epithelial cell line
was able to generate itself as well as MUC+/ESA+
epithelial cells and Thy-1+/
-smooth muscle
actin+ (ASMA+) myoepithelial cells. The
MUC/ESA+ epithelial cell line further differed
from the MUC+/ESA+ epithelial cell line by the
expression of keratin K19, a feature of a subpopulation of epithelial
cells in terminal duct lobular units in vivo. Within a reconstituted
basement membrane, the MUC+/ESA+ epithelial cell
line formed acinus-like spheres. In contrast, the
MUC/ESA+ epithelial cell line formed elaborate
branching structures resembling uncultured terminal duct lobular units
both by morphology and marker expression. Similar structures were
obtained by inoculating the extracellular matrix-embedded cells
subcutaneously in nude mice. Thus, MUC/ESA+
epithelial cells within the luminal epithelial lineage may function as
precursor cells of terminal duct lobular units in the human breast.
[Key Words: Human cell; breast; stem cell; immortal; K19]
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