Effects of growth factors on proliferation on basal and luminal cells in human breast epithelial explants in serum-free culture.
A method of culturing human breast epithelium is described in which viable explants can be maintained in protein-free medium while retaining the capacity of responding to added hormones and growth factors for at least 7 days. Culture parameters were chosen to provide maximum sensitivity of detection of proliferative responses by autoradiography. Under basal conditions, the mean thymidine labeling index of the explants was 0.08%. After stimulation with insulin, hydrocortisone, and cholera toxin (I,H,CT), a combination known to stimulate proliferation in human breast epithelium in vitro, the mean labeling index was 15.7%. Stimulation of explants with epidermal growth factor (EGF) and transforming growth factor (TGF)-alpha resulted in mean labeling indices of 6.6 and 10.8%, respectively. Autoradiography at the ultrastructural level demonstrated that in I,H,CT-stimulated explants the majority of the labeled cells were luminal, with only 1.5% being basal cells. In contrast, after EGF and TGF-alpha basal cells accounted for 11.5 and 18.5% of the labeled population. These results indicate that this system provides an in vitro assay of proliferative activity in the normal human breast that enables comparisons to be made between both the luminal and the basal cells in the explants and their counterparts in monolayer culture prepared from flow sorted cells. Thus, growth responses dependent on cell-to-cell interactions or stromal modulation can be identified.
Perusinghe
N P
NP
Institute of Cancer Research, Haddow Laboratories, Sutton, Surrey, United Kingdom.
Monaghan
P
P
O'Hare
M J
MJ
Ashley
S
S
Gusterson
B A
BA
D001940:Breast / Q000166:cytology*; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D004247:DNA / Q000096:biosynthesis; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D004847:Epithelial Cells; D006801:Humans; D008854:Microscopy, Electron; D013997:Time Factors; D016211:Transforming Growth Factor alpha / Q000494:pharmacology*
D016895:Culture Media, Serum-Free; D016211:Transforming Growth Factor alpha; D004815:Epidermal Growth Factor; D004247:DNA