Figure 1 Primary cell culture of ovary cancer tumour.
Figure 2 Follow up of cell viability for representative samples 0, 5 and 10 days after seeding.
Figure 3 Representative example of correlation between primary tumour histology and cell culture (morphology and immunophenotype) from an endometrioïd ovarian adenocarcinoma grade 2. Cell culture and, within the box, histology of the tumour are stained by haematoxilin and eosin. Progesterone receptor (PR) immunostaining for ovarian cultured cells and primitive tumour tissue (box) were observed. WT-1 labelings in the cultured cells and primitive tumour tissue (box) are negative. Negative immunostaining for CD163 and calretinin excluded the macrophagic or mesothelial nature of these cells.
Figure 4 Rate of cell death in control and treated cells for patients A, B, C and D. Treated cells were exposed for 72 hours to carboplatin (Carbop.), vepeside (Vep.), topotecan (Topot.) and paclitaxel (Pacl.).
Figure 5 Statistical analyses of cell death rate after treatment with carboplatin, vepeside, topotecan or paclitaxel.
Antibody |
Supplier |
Clone |
Dilution |
Antigen retrieval / Incubation time |
Vimentin |
DAKO, Glostrup, Denmark |
V9 |
1/200 |
CC1 mild/20’ |
WT-1 |
DAKO |
6F-H2 |
1/50 |
CC1 standard/32’+ amplif |
PR |
Ventana, Rocklin, CA, USA |
1E2 |
prediluted |
CC1 standard/32’ |
ER |
Ventana |
SP1 |
prediluted |
CC1 standard/32’ |
P53 |
DAKO |
D07 |
1/50 |
CC1 standard/32’ |
P16 |
Ventana |
inK4a |
prediluted |
CC1 standard/32’ |
HNF1beta |
Atlas Antibodies, Stockholm, Sweden |
poly |
1/200 |
CC1 standard/60’+ amplif |
CD163 |
Novocastra, Newcastle upon Tyne, UK |
10D6 |
1/100 |
CC1 mild/32’ |
CD68 |
DAKO, Glostrup, Denmark |
PGM1 |
1/50 |
CC1 mild/20’ |
Calretinin |
ZYMED, San Francisco, CA |
poly |
1/100 |
CC1 mild/32’ |
Table1 Antibody list and staining procedures.
Patient |
Pathology |
Culture success |
1 |
low grade serous adenocarcinoma (type 1) |
yes |
2 |
high grade serous adenocarcinoma (type 2) postchemotherapy |
Contamination |
3 |
metastatic ductal breast carcinoma |
yes |
4 |
endometrioid adenocarcinoma grade 2 FIGO |
yes |
5 |
clear cell adenocarcinoma |
yes |
6 |
clear cell adenocarcinoma |
yes |
7 |
high grade serous adenocarcinoma (type 2) |
yes |
8 |
serous borderline cystadenoma |
yes |
9 |
high grade serous adenocarcinoma (type 2) |
yes |
10 |
high grade serous adenocarcinoma (type 2) |
yes |
11 |
high grade serous adenocarcinoma (type 2) postchemotherapy |
yes |
12 |
clear cell adenocarcinoma |
contamination |
13 |
serous-transitional cell carcinoma |
yes |
14 |
endometrioid adenocarcinoma grade 2 FIGO |
yes |
15 |
endometrioid adenocarcinoma grade 2 FIGO |
yes |
16 |
clear cell adenocarcinoma |
yes |
17 |
clear cell adenocarcinoma |
yes |
18 |
high grade serous adenocarcinoma (type 2) |
yes |
19 |
mature teratoma |
yes |
20 |
high grade serous adenocarcinoma (type 2) |
yes |
21 |
endometrioid adenocarcinoma grade 2 FIGO |
yes |
22 |
endometrioid adenocarcinoma grade 2 FIGO |
yes |
23 |
mucinous borderline cystadenoma |
yes |
24 |
dysgerminoma |
yes |
25 |
high grade serous adenocarcinoma (type 2) |
yes |
26 |
high grade serous adenocarcinoma (type 2) |
yes |
27 |
high grade serous adenocarcinoma (type 2) postchemotherapy |
yes |
28 |
high grade serous adenocarcinoma (type 2) and low grade serous adenocarcinoma (type 1) |
yes |
Table2 Histological diagnoses of all 28 samples and successes in culture.
Case |
Diagnosis |
WT1 |
VIM |
RE |
RP |
P16 |
P53 |
HNF- |
8 |
serous borderline cystadenoma |
+ |
+ |
+ |
ND |
ND |
ND |
ND |
10 |
high grade serous adenocarcinoma (type 2) |
+ |
+ |
+ |
ND |
ND |
ND |
ND |
16 |
clear cell adenocarcinoma |
ND |
ND |
ND |
ND |
ND |
ND |
+ |
18 |
high grade serous adenocarcinoma (type 2) |
+ |
- |
- |
ND |
+ |
- |
ND |
19 |
mature teratoma |
ND |
ND |
ND |
ND |
ND |
ND |
ND |
21 |
endometrioid adenocarcinoma grade 2 FIGO |
- |
- |
+ |
+ |
+ |
ND |
- |
25 |
high grade serous adenocarcinoma (type 2) |
+ |
- |
- |
+ |
NC |
- |
ND |
27 |
high grade serous adenocarcinoma (type 2) |
- |
- |
+ |
ND |
+ |
+ |
ND |
28 |
high grade serous adenocarcinoma (type 2) |
+ |
- |
+ |
- |
- |
ND |
- |
Table3 List of the 9 interpretable cases. Immunocytochemical features. (ND: not determined)
Patient |
Pathology |
Proliferation (%) |
17 |
clear cell adenocarcinoma |
65,6 |
21 |
endometrioid adenocarcinoma grade 2 FIGO |
23,7 |
23 |
mucinous borderline cystadenoma |
21,7 |
24 |
dysgerminoma |
18,3 |
25 |
high grade serous adenocarcinoma (type 2) |
16,3 |
27 |
high grade serous adenocarcinoma (type 2) |
19,5 |
Table4 Percentage of proliferative cells for 6 representative cultures determined by BrdU incorporation.