Introduction
Methods
Clinical specimens of malignant PT
Animal model
Immunohistochemical analysis
Immunofluorescent staining
Cell sorting and analysis by flow cytometry
Mammosphere assay
Differentiation of human tumor stem cells
Special stains
Results
Successful engraftment of primary human PTs in NOD-SCID mice
Phenotypic markers of primary PTs and xenografts
% positive cases (n = 51) | BC007/BC-P007MT1 | BC107/BC-P107MT1 | BC515/BC-P515MT1 | BC877/BC-P877MT1 | HS-5 | ||
---|---|---|---|---|---|---|---|
(% positive cells) | |||||||
MSC marker | CD44 | 68.6% | 50/60 | 60/30 | -/- | -/- | + |
CD29 | 27.4% | 10/40 | <5/30 | 5/<1 | -/- | + | |
CD166 | 78.4% | <5/< 5 | 10/50 | 5/5 | 90/90 | + | |
CDI06 | 29.4% | 30/<10 | <10/<10 | -/- | 80/80 | + | |
CD105 | 1.9% | -/- | -/- | -/- | 50/90 | + | |
CD90 | 78.4% | <10/< 10 | 10/10 | 55/75 | <5/70 | + | |
CD117 | 70.5% | -/20 | -/- | 5/50 | 80/95 | + | |
GD2 | 100% | 80/50 | 75/30 | 90/75 | 60/30 | + | |
(<10 to >90) | |||||||
Stem cell marker | ALDH | 100% | 20/1 | 3-5/<1 | 40/5 | <1/<5 | + |
(<10 to >90) | |||||||
Oct-4 | 47.0% | 10/5 | 30/10 | <1/35 | <5/<5 | + | |
PT clinically relevant markers | Vimentin | 100% | 90/ > 90 | 90/>70 | 80/20 | >95/>95 | + |
(<10- to >90) | |||||||
Ki-67 | 82.3% | 40/40 | 10/20 | 5/<3 | 65/>95 | + | |
CD10 | 60.7% | 20/5 | 10/70 | 5/5 | <10/30 | + | |
CD34 | 52.9% | -/- | 40/70 | 80/20 | -/- | - | |
p53 | 50.9% | <10/<10 | <5/10 | 70/60 | >95/>95 | + | |
Bcl-2 | 37.2% | -/- | -/- | -/- | -/- | - | |
p63 | 9.8% | 60/10 | -/- | -/- | -/- | - | |
Globo-H | 9.8% | -/- | -/- | -/- | 80/80 | - |
ALDH+identifies cancer stem cells in malignant PTs
BC-P007 | 4 × 104 | 2 × 104 | 1 × 104 | 5 × 103 | 4 × 103 | 1 × 103 | 500 | 100 | 50 | 20 | Frequency〒 | Distribution# |
---|---|---|---|---|---|---|---|---|---|---|---|---|
ALDH+ | 3/8 | 3/8 | 1/3 | 1/7 | 1:2.0 × 104 | 75% | ||||||
ALDH- | 0/4 | 0/6 | 1/7 | 0/3 | 0/4 | 1:1.1 × 105 | 25% | |||||
ALDH+/GD2+ | 2/2 | 3/3 | 6/10 | 5/10 | 2/20 | 0/3 | 1:6.1 × 102 | |||||
ALDH-/GD2- | 0/4 | 0/1 | 0/8 | 2/10 | 0/15 | 1:1.3 × 105 | ||||||
ALDH+ | 3/8 | 3/8 | 1/3 | 1/7 | 1:2.0 × 104 | |||||||
ALDH+/GD2+ | 2/2 | 3/3 | 6/10 | 5/10 | 2/20 | 0/3 | 1:6.1 × 102 | |||||
BC-P515
|
1 × 10
5
|
2 × 10
4
|
1 × 10
4
|
4 × 10
3
|
1 × 10
3
|
500
|
100
|
50
|
Frequency
〒
|
Distribution
#
| ||
ALDH+ | 1/4 | 0/3 | 1/4 | 3/9 | 1/5 | 1:2.0 × 104 | 96% | |||||
ALDH- | 0/4 | 0/3 | 0/2 | 0/9 | 1:>4.2 × 105 | 4% | ||||||
ALDH+/GD2+ | 1/3* | 7/10 | 4/5 | 0/1 | 2/7 | 1/2 | 1:7.1 × 103 | |||||
ALDH-/GD2- | 0/1 | 0/2 | 1/9 | 0/4 | 0/10 | 1:2.5 × 105 | ||||||
ALDH+ | 1/4 | 0/3 | 1/4 | 3/9 | 1/5 | 1:2.0 × 104 | ||||||
ALDH+/GD2+ | 1/3 | 7/10 | 4/5 | 0/1 | 2/7 | 1/2 | 1:7.1 × 103 |