Introduction
Material and methods
Cell culture
Lentivirus infections
Ex vivoniche cultures
Dissociation of ex vivoniche cultures
Antibodies and immunochemistry
Microscopy and image analysis
hAMSCs differentiation assays
Flow cytometry
Microarray profiling
Immunoblotting
Statistical analysis
Results
Isolation and propagation of human breast epithelial and mesenchymal precursor cells
PHBECs and hAMSCs maintain their identity when grown on nylon meshes submerged in M5 medium
Breast epithelial and mesenchymal precursors constrain growth and spreading reciprocally in heterotypic cultures
Ex vivoheterotypic culture conditions maintain mesenchymal and breast epithelial precursor cells
Transforming growth factor (TGF)β signaling is active in mesenchymal cells from co-cultures
Upstream regulator | Predicted activation state | Activation z-score | P-value of overlap |
---|---|---|---|
BNIP3L | Activated | 3.05 | 5.60E-12 |
TP53 | Activated | 2.82 | 8.44E-12 |
let-7 | Activated | 2.01 | 1.12E-10 |
KDM5B | Activated | 2.90 | 7.65E-07 |
CDKN2A | Activated | 2.70 | 1.91E-06 |
MAPK14 | Activated | 2.28 | 5.65E-06 |
TCF3 | Activated | 3.32 | 1.49E-05 |
CEBPD | Activated | 2.20 | 1.25E-04 |
PTHLH | Activated | 2.20 | 1.47E-04 |
UXT | Activated | 2.22 | 1.53E-04 |
TAZ | Activated | 2.00 | 6.60E-04 |
MAP3K7 | Activated | 2.30 | 7.45E-04 |
SMARCB1 | Activated | 2.14 | 1.47E-03 |
P38 MAPK | Activated | 2.29 | 6.01E-03 |
JUN | Activated | 2.03 | 1.85E-02 |
Vegf | Inhibited | −2.35 | 1.70E-09 |
CCND1 | Inhibited | −2.24 | 1.12E-08 |
MYC | Inhibited | −4.12 | 1.20E-08 |
E2F1 | Inhibited | −2.89 | 2.51E-07 |
TBX2 | Inhibited | −2.53 | 8.13E-07 |
FOXO1 | Inhibited | −3.56 | 1.67E-05 |
MYBL2 | Inhibited | −2.00 | 3.49E-03 |
MYCN | Inhibited | −2.38 | 7.17E-03 |
BMP7 | Inhibited | −2.39 | 1.19E-02 |
CDKN1A | 1.60 | 4.35E-21 | |
HGF | −1.77 | 1.39E-10 | |
CDK4 | 1.82E-09 | ||
TGFβ1 | 0.68 | 9.63E-08 | |
RB1 | 1.14 | 8.23E-07 | |
Tgf β | 0.88 | 9.33E-06 |
Upstream regulator | Predicted activation state | Activation z-score | P-value of overlap |
---|---|---|---|
TGFβ1 | Activated | 2,580 | 4,76E-16 |
SMAD2 | Activated | 2,204 | 7,91E-06 |
SMAD3 | Activated | 2,138 | 8,05E-06 |
NOTCH1 | Activated | 2,102 | 3,87E-05 |
Tgf β | Activated | 2,241 | 5,70E-05 |
MKL1 | Activated | 2,744 | 4,28E-04 |
MKL2 | Activated | 2,449 | 2,49E-03 |
MGEA5 | Activated | 2,887 | 6,60E-03 |
INSIG1 | Activated | 2,621 | 9,33E-03 |
GFI1 | Activated | 2,236 | 1,56E-02 |
TNF | Inhibited | −2,756 | 1,52E-15 |
IL1B | Inhibited | −2,446 | 2,96E-09 |
TP53 | Inhibited | −2,871 | 5,83E-08 |
NFkβ (complex) | Inhibited | −2,030 | 1,20E-07 |
CEBPA | Inhibited | −2,457 | 1,66E-06 |
TP63 | Inhibited | −2,301 | 5,32E-06 |
RELA | Inhibited | −2,192 | 5,25E-05 |
PDGF BB | Inhibited | −2,883 | 2,02E-04 |
MYCN | Inhibited | −2,440 | 9,87E-04 |
ALDH2 | Inhibited | −2,000 | 1,17E-03 |
CHUK | Inhibited | −2,101 | 1,39E-03 |
PPARG | Inhibited | −2,476 | 1,46E-03 |
TGFB3 | 1,286 | 4,60E-08 | |
INHBA | −0.341 | 7.33E-05 | |
TGFβR2 | −0,296 | 1,59E-04 | |
SMAD4 | 0,659 | 1,80E-04 | |
BMP7 | −0,464 | 2,17E-04 |